Stem Cells · 16 Father of Hybrid Rice Conferred 2011 Mahathir Science Award 16 Personalized...

Stem Cells

Expatriate Researcher Returns to Roost

Past, Present and Future of Stem Cells in Regenerative Medicine

Learning Human Cardiac Diseases through Stem Cells

Bioprocessing for Stem Cell Therapy: From the Lab into the Log Phase

Neural Stem Cells and Cell-based Approaches in Neurodegeneration and Peripheral Nerve Injuries

The Marketing of Unapproved Stem Cell Products: An Industry-wide Challenge




Bioprocessing for Stem Cell Therapy: From the Lab into the Log Phase

Neural Stem Cells and Cell-based Approaches in Neurodegeneration and Peripheral Nerve Injuries


March 2012 Vol. 16 • No. 3 A monthly international biotechnology publication

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CONTENTSEDITORIAL

BIOBOARD

AUSTRALIA

5 Aussie Meteorite Yields Anti-Epileptic Amino Acid

6 Novel Partnership Could Transform Breast Cancer Diagnosis

6 University of Sydney Academics Laud Sunbed Ban

7 Australian Universities Losing Appeal to Asians

INDIA

7 New Nanobiotechnology Lab in Gurgaon

8 China, India, and Myanmar Collaborate to Develop Brahmaputra-Salween Ecosystem

9 Incredible India – Now Polio-Free

9 Increase Women’s Participation in Science for Over-All Development of Nation

JAPAN

10 DNA Motor Now Can Navigate Networks

10 RIKEN’s Diagnosis Technique Detects H1N1 at Record Speed

11 New Hotspot on Rice Chromosome Could Yield Improved Varieties

OTHER REGIONS

11 Delivering RNAi the Spongy Way

EYE ON CHINA 12 China Unveils World-Class Biotech Ambitions

13 Chinese Scientists Awarded Cancer Research Prize

14 BGI Researchers Uncover Extensive RNA Editing in Human Transcriptome

15 Johns Hopkins Commences New China STEM Program

16 Father of Hybrid Rice Conferred 2011 Mahathir Science Award

16 Personalized Medicine Receives Boost in China

17 GeneNews & Shanghai Biochip to Establish First Sentinel Centre for Personalized Medicine

18 SINANO Invents New Microfluidic Chip for Bacterial Culture

Upcoming IssuesAprilBioEthics

MayBiofuels

JuneClinical Research Labs in China

FEATURES20 Expatriate Researcher Returns to Roost

24 Past, Present and Future of Stem Cells in Regenerative Medicine

28 Learning Human Cardiac Diseases through Stem Cells

32 Bioprocessing for Stem Cell Therapy: From the Lab into the Log Phase

36 Neural Stem Cells and Cell-based Approaches in Neurodegeneration and Peripheral Nerve Injuries

43 The Marketing of Unapproved Stem Cell Products: An Industry-wide Challenge

REVIEW47 Annual Pivotal Event for Global Chinese Medicine Industry

INSIDE INDUSTRY48 India Unveils Roadmap for Cooperation with Africa

49 Indian Agriculture Minister Calls For Improved Soil Health

50 A*STAR, GE Global Research To Develop Integrated Advanced Medical Imaging

51 PRA Opens Office in Singapore

52 Bristol-Myers Squibb Announces Grants to Tackle Hepatitis B and Hepatitis C in Asia

53 Burrill & Company Expands into Taiwan

53 Medistem Initiates Collaboration With Shanghai Jia Fu Medical Apparatus

54 Advinus, SignalChem Commence Collaboration On New Anti-Cancer Drugs

55 3D Petri Dish to Reduce Animal Use in Research

55 Vermillion Receives Patent on Platelet Biomarkers of Angiogenesis

56 Biotech R&D Outsourcing Set to Increase and Shift Overseas

CONFERENCE CALENDAR

3

EDITORIALStem cells have often been dubbed the “fountain of youth”, due to their amazing regenerative capabilities. But while we are still far away from any such elixir, there is no denying their immense potential in treating human diseases. In this special issue on stem cells, we have gathered a good mix of articles for scientists, industrialists and laymen alike. We trace the past, present, and future of stem cells in regenerative medicine, uncovering little-known nuggets of trivia that may surprise and delight even hardcore researchers. We describe cutting-edge approaches to treating neurodegeneration and nerve injuries too. None of these therapies would work however if we are unable to produce stem cells on a massive scale for animal studies, clinical trials and treatment. Enter, the new age of bioprocessing techniques devised by the Bioprocessing Technology Institute in Singapore, promising to grow billions to trillions of cells consistently for a variety of purposes. Treating diseases though, is just one of stem cells’ myriad applications. They can be used to model diseases too, as researchers at National Heart Centre Singapore have shown. Given all the pressure to move stem cells from bench-to-bedside, we should not be surprised at the proliferation of certain dubious treatments slipping through the cracks of our drug regulation systems. Here, we shed light on the murky world of unapproved stem cell products – a phenomenon that speaks volumes of the huge promise that stem cells hold in the minds of consumers.At APBN, we believe that stem cells will enjoy many more years in the limelight. There remain several challenges to their incorporation into mainstream medicine; in spite of this, we look forward to new discoveries in the field that uncover hitherto unknown vast reserves of therapeutic and biotechnological potential.

V. K. SanjeedEditor, Asia Pacific Biotech News

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BIOBOARD

AUSTRALIA

Aussie Meteorite Yields Anti-Epileptic Amino Acid

D ecades after it crashed into a field in rural Australia, the Murchison meteorite remains one of the most

studied meteorites due to its richness in organic compounds, including amino acids. Scientists at Albany Medical College have found that one such extraterrestrial amino acid may prevent or reduce the duration of seizures in epilepsy. Their findings were published in the journal Epilepsia.

The presence of amino acids—the molecules that make up proteins, and hence essential to life as we know it—within an extra-terrestrial object has led scientists to question whether the meteorite is a clue to life outside of our solar system, and whether theories of evolution’s “big bang theory” hold any truth. Among the amino acids found within the meteorite, of particular interest to Damian Shin, Ph.D., Assistant Professor

in the Center for Neuropharmacology and Neuroscience at Albany Med, is the amino acid, isovaline.

Prior to this, isovaline has been found to be structurally similar to glycine, a more down-to-earth amino acid commonly found in proteins here on Earth. Glycine is known to calm nerves in the brain so Dr Shin hypothesized that its extra-terrestrial counterpart may also have an effect on seizure control.

His intuition paid off. Using rodent models, Dr. Shin and his team found that isovaline quiets excitatory neurons, albeit via a mechanism unlike glycine’s. By increasing the activity of interneurons, which act as ‘gating’ cells, which control how other neurons interact with each other, isovaline stopped seizures completely or reduced the frequency of seizures by 50 percent.

“This is promising because we know that epilepsy causes a disruption in the normal functioning of some ion channels—prominent components of the nervous system that conduct nerve impulses within a cell. Sometimes the activity of ion channels become pathological, which makes some neurons ‘hyperexcited’,” says Dr. Shin. “The next step is to determine which ion channel is affected by this amino acid. This will help us better understand how isovaline can impart anticonvulsant properties for patients with epilepsy.”

He stresses that the work is preliminary and it could be years before the work is translated to a treatment for humans. However, he is encouraged by these early results to possibly help a portion of the population who do not respond to normal epilepsy drugs or experience serious side effects.

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BIOBOARD

A n innovative University of Sydney research partnership with private enterprise could see a radical change

in how breast cancer is diagnosed, with the possibility to improve detection rates.

The partnership is utilising advanced technology made available by Agilent Technologies, a company using novel imaging tools such as elemental bio imaging and microwave imaging, to allow researchers from the University’s Charles Perkins Centre and Faculty of Health Sciences to study breast tissue samples in an entirely different way.

The project is the first study of its kind to look at measuring physical properties of breast tissue which could be related to breast cancer risk. Breast tissue density has been widely recognised as being strongly linked to the risk of developing breast cancer. Currently, the most common approach to determining a woman’s breast density is for a radiologist to visually assess the percentage of fibroglandular tissue imaged on a mammogram, and assign it to a category of density from 1-4, which is known as the Breast Imaging Reporting and Data System (BI-RADS) classification system.

Project lead Dr Elaine Ryan said a more accurate and reliable measurement of breast density was urgently required as the traditional technique using mammogram imaging is inaccurate, subjective and unreliable.

“This is due to the measurement of a 3D volume from a 2D representation of the tissue. In addition, mammograms are not being used to help determine any breast cancer risk models, despite the fact it is widely recognised that analysis of breast density data would be a critically important addition to ways of diagnosing the condition,” she said.

Dr Ryan said the research project would determine whether measuring the structure and composition of breast tissue, using x-ray fluorescence signals, could be used to accurately measure breast density and predict risk of developing the condition.

“A more accurate measure - that does not depend on the subjective opinion of radiologists - would improve the reliability of breast density estimates for risk prediction and could lead to improved targeted screening programs for high risk women,” she said.

The novel experimental approach will combine a measurement of the electron density, trace element content, liquid content and fat cell content, into a model which can be correlated with mammographic breast density measurements.

Dr Ryan said the research team believed this approach could show that glandular tissue structure and composition can provide a direct measure of breast tissue density.

“This in turn has the potential to be able to develop more accurate methods of calculating breast density from conventional mammographic imaging. By developing a better understanding of the structure of dense tissue, better models can be developed which consider a more complex structure of tissue, rather than the bi-tissue type assumption currently used.

An accurate measure of breast density will vastly improve current risk models, allowing better decisions to be made about an individual’s screening program, and whether preventative measures may be appropriate to manage women’s risk of developing this disease.

Novel Partnership Could Transform Breast Cancer Diagnosis

A NSW government decision to ban solariums has been welcomed by University of Sydney researchers

whose study contributed to the move.“This decision by the NSW government

is groundbreaking. NSW is the first state in Australia to propose such legislation and Australia will be only the second country in the world, after Brazil, to take this step,” said Professor Bruce Armstrong of the Sydney Medical School, who in 2011 co-authored the Australian Melanoma Family Study (AMFS) report.

The study, published in the International Journal of Cancer, was the first to examine the association of sunbed use with melanoma occurrence in younger adults. It found that the use of sunbeds by young people aged 18 to 39 years increased their risk of developing melanoma by an average of 41 percent.

University of Sydney Academics Laud Sunbed BanAnother researcher, Professor Graham

Mann, who leads melanoma research at the Westmead Millennium Institute for Medical Research, was the lead author on the only Australian study into increased melanoma incidence and use of sun beds by young people. His study showed that the risk for people under 30 was, in fact, even higher.

“For people who got melanoma under the age of 30, and who had used sunbeds at all, 75 per cent of those melanomas are attributable to sunbeds. We are delighted that our research has contributed to a very important health measure that will help prevent melanoma in New South Wales, and hopefully, if other jurisdictions pick it up, will save more lives around Australia,” said Professor Mann.

The Australian Melanoma Family Study findings were the result of a research

collaboration between the University of Sydney, Westmead Millennium Institute for Medical Research, University of Melbourne, Melanoma Institute Australia, Cancer Council Victoria and Cancer Council Queensland.

Skin cancers account for 80% of all newly diagnosed cancers in Australia. Skin cancer is also the most expensive cancer. In 2001, it was estimated the treatment of non-melanoma skin cancer cost $264 million and melanoma $30 million.

7

BIOBOARD

F or years, Australian universities have enjoyed a steady flow of students from all parts of Asia seeking to undertake

undergraduate and postgraduate studies. The country’s relative geographical proximity to students from India, China and Southeast Asia, its world-class infrastructure, not to mention top-notch university faculties that offered the promise of a “Western” brand of higher education, were all factors that worked to Australia’s favour.

That favour, is now being withdrawn, according to Professor Greg McCarthy, Head of the School of Social Sciences at the University of Adelaide. Speaking at The Future of Higher Education Conference recently, the expert on Asian education pointed out that Australian schools and universities had neglected Asian studies – and exploited international students – for so long that “they would really struggle to

Australian Universities Losing Appeal to Asiansfind a way back from here”.

The problem is most manifest in the dearth of Asian studies departments in universities. In 2009, just 300 students from non-Chinese backgrounds studied Mandarin at year 12 level. Indonesian language studies have fared no better. The situation reflects an educational policy that seems outdated and out of touch with the realities of the Asian century.

A lack of Asian-focused studies however, is just the tip of the iceberg. In 2009, a series of racially motivated crimes against Indians in Australia and a large protest organised by Indian students brought Australia into the spotlight and cast a pall over India – Australia relations. Indian students are the second largest group of international students studying at a tertiary level in Australia, and their safety is an issue of concern not just to their families, but

the Indian government as well. In spite of numerous official investigations into the attacks, and efforts to improve the lot of Indian students, the damage was done. The reputation of Australia as a prime destination for higher education has since suffered on the Sub-continent, and students increasingly look towards other countries like USA for further studies.

Aust ra l ia needs to re look i t s educational policies if it wants to keep its slice of the highly lucrative Asian tertiary education market. Its universities can take the first crucial steps by ensuring international students engage with their local counterparts, experience the culture and speak the language. But perhaps even more crucial, is the need for Australia to rethink its position in Asia and prepare for more competition from Asian universities

D eakin University and The Energy and Resources Institute (TERI) have opened a state-of-the-art

‘Nanobiotechnology Research Centre’ in Gurgaon, India, dedicated to researching solutions for a greener and more advanced use of nanotechnology for resolving challenging agricultural, biomedical and sustainability issues.

The centre’s development was an outcome of TERI’s core capability of knowledge creation and development of efficient, environment friendly technologies and Deakin’s India Research Initiative (DIRI) which is committed towards establishing a lasting association with industry partners in India to chart a vibrant culture of research and scholastic excellence.

INDIA

New Nanobiotechnology Lab in Gurgaon

Hon’ble Louise Asher, MP and Minister for Innovation, Services & Small business, Minister for Tourism and Major Events, Australia along with Professor Jane den

Hollander, Vice Chancellor, Deakin University and Dr RK Pachauri, Director General TERI inaugurated the facility on 22nd February 2012. The new laboratory will be used to bring together Deakin University’s expertise in the design and characterisation of novel nanomaterials and TERI’s experience in biotech applications in food, agriculture, environment and pharmacology.

Interest in using bionanotechnology to help a country’s food security issue, provide benefits in the area of health as well as address environmental issues, has been gaining momentum in India as well as in Australia in recent years. The centre aims to house about 70 researchers including 50 PhD students enrolled at Deakin and co-supervised by Deakin and TERI practitioners.

8

BIOBOARD

R epresentatives from China, India, and Myanmar gathered in Myanmar from 21 to 23 December to plan the

transboundary management of a biologically rich Himalayan ecosystem shared by the three countries. The programme framework for a regional Brahmaputra-Salween landscape initiative was formulated at an expert consultation held in Nay Pyi Taw, organised jointly by the Ministry of Environmental Conservation and Forestry, Government of the Republic of Myanmar, and the International Centre for Integrated Mountain Development (ICIMOD).

The Brahmaputra-Salween landscape comprises several remote but key protected areas in the eastern Himalayas, including Gaoligongshan National Nature Reserve in China, Namdapha National Park in India (also a tiger reserve), and Hkakaborazi National Park in Myanmar. The area is important not only from the national perspectives of the

China, India, and Myanmar Collaborate to Develop Brahmaputra-Salween Ecosystem

participating countries, but also globally; it is home to a number of wildlife species of global importance such as takin, red panda, snub nosed monkey, hollock gibbon, and Namdapha flying squirrel, as well as many endemic flowering plants. These globally important species are distributed widely across the landscape, irrespective of the national boundaries. Therefore, noted Dr David Molden, Director General of ICIMOD, “a regional approach is required to manage this mountain landscape, to enhance the livelihoods of the people living there, and to conserve its natural resources and ecosystem services for future generations”.

The consultation to develop a future strategic programme for transboundary biodiversity management and climate change adaptation in the Brahmaputra-Salween landscape was inaugurated by His Excellency U Win Tun, Minister for Environmental Conservation and Forestry, Myanmar, who

urged the participating countries to focus on both research and development to take this initiative further. Other speakers included U Nyi Nyi Khaw, Deputy Director, Department of Forest, Myanmar and Chair of the Board of Governors, ICIMOD; Dr Li Dezhu, Director of the Kunming Institute of Botany, China; Dr LMS Palni, Director of the GB Pant Institute of Himalayan Environment and Development, India; and Dr Eklabya Sharma, Director of Programme Operations, ICIMOD.

The 30 part ic ipants drafted a programme framework whose highlights include collaborative and multidisciplinary research, regional capacity building, and policy and institutional support. Planned interventions will promote transboundary b iodivers i ty management , cultura l conservation, sustainable economic development, and enhanced ecosystem and socio-economic resilience in the Brahmaputra-Salween landscape.

9

BIOBOARD

O n the 25th of February, the World Health Organization notified Indian authorities that the country was

officially removed from the list of countries with active transmission of endemic polio. The announcement was made after India successfully marked 12 months in which no Indian child had been paralyzed by polio. WHO data showed that samples from the previous year cleared laboratory tests with no sign of wild poliovirus, indicating that for the first time in history, there is no polio in India.

The official announcement was made at the Polio Summit 2012 in New Delhi. A letter written by WHO Director-General Dr Margaret Chan and delivered to the Indian Government on the morning of the Polio

Incredible India – Now Polio-FreeSummit confirmed the news. The momentous occasion was greeted with thundering applause and a standing ovation by the 1400-plus participants from across the world. India’s Health Minister, Ghulam Nabi Azad, said: “We have won the battle but the war is not yet over. Let us today rededicate ourselves and resolve that we will continue our efforts with the same vigour, so that India can be declared (certified) polio-free by 2014.”

Prime Minister Manmohan Singh said that the success of India’s efforts was proof that teamwork pays, adding that the Union and State Governments have worked in close partnership with many community, national and international organizations and partners including Rotary International, World Health Organization, UNICEF and the US Centers for

Disease Control and Prevention to eradicate polio.

Still, the Health Minister assured the audience that the Indian Government was “acutely aware” that it could not rest on its laurels. “We are excited and hopeful, at the same time, vigilant and alert. We are highly mindful of the risks that persist, not only on account of residual indigenous transmission but also from other countries.” Stressing that “there is no room for complacency”, Mr Azad said the programme needs to continue with “full force” until polio was eradicated globally.

India’s success leaves only three countries remaining polio-endemic - meaning they have never stopped indigenous wild poliovirus transmission: Afghanistan, Nigeria and Pakistan.

S m t N i r u p a m a R a o , I n d i a n Ambassador to the US, has called f o r i n c r e a s e d p a r t i c i p a t i o n

and retention of women scientists for overall development of the nation. Inaugurating the first Women Science Congress on the sidelines of the ongoing 99th Indian Science Congress (ISC) at Bhubaneshwar, Smt Rao called for redoubling efforts to make science more inclusive. She said that we need to encourage the participation of women in science in India. The former foreign secretary called for policy solutions to remove gender-based disparity in science and other sectors.

"We have to encourage participation and retention of Indian women scientists in science and technology, which is important for national development and the success of other programmes," she added.

The Jan 3-7 event on the theme 'Science and Technology for Inclusive Innovation - Role of Women' is focused on women in science. In line with the theme, the

Increase Women’s Participation in Science for Over-All Development of Nation

five-day ISC was headed by Geetha Bali, vice chancellor of the Karnataka State Women's University, Bijapur. She is the fourth woman in the history of the ISC to head the Congress.

Addressing the Women Science Congress, D. Purandeswari, Minister of State for Human Resource Development (HRD), said the development of women cannot be complete without their equal participation in science. Earlier, inaugurating the ISC Tuesday, Prime Minister Manmohan Singh underlined the need for transparency in selection procedures and also the importance of gender audits.

"We should also take note of the results of a study published last year that showed that 60 percent of nearly 2,000 Indian women PhDs in science who were surveyed were unemployed. The main reason cited was lack of job opportunities. Only a very small number cited family reasons. This underlines the need for transparency in selection procedures at institutions and also the importance of gender audits," she said.

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BIOBOARD

E xpanding on previous work with engines traveling on straight tracks, a team of researchers at Kyoto

University and the University of Oxford have successfully used DNA building blocks to construct a motor capable of navigating a programmable network of tracks with multiple switches. The findings, published in the January 22 online edition of the journal Nature Nanotechnology, are expected to lead to further developments in the field of nanoengineering.

The research utilizes the technology of DNA origami, where strands of DNA molecules are sequenced in a way that will cause them to self-assemble into desired 2D

JAPAN

DNA Motor Now Can Navigate Networksand even 3D structures. In this latest effort, the scientists built a network of tracks and switches atop DNA origami tiles, which made it possible for motor molecules to travel along these rail systems.

“We have demonstrated that it is not only possible to build nanoscale devices that function autonomously,” explained Dr. Masayuki Endo of Kyoto University’s Institute for Integrated Cell-Material Sciences (iCeMS), “but that we can cause such devices to produce predictable outputs based on different, controllable starting conditions.”

The team, including lead author Dr. Shelley Wickham at Oxford, expects that the work may lead to the development

A new diagnosis technique developed by researchers at the RIKEN Omics Science Center (OSC) has succeeded

in detecting influenza virus infection in only 40 minutes and with one hundred times the sensitivity of conventional methods. Clinical research conducted in 2009 and 2010 confirms the new technique accurately identified the 2009 pandemic (pdm) influenza virus in Japanese patients less than 24 hours after fever onset, much faster than standard diagnostic tests.

The human-to-human transmission of new, highly pathogenic strains of influenza virus poses today a major threat to human health and to the security of global society. With its rapid global spread, the 2009 pandemic (pdm) influenza virus reminded the world of this threat, resulting in an estimated 18,000 deaths worldwide. In Japan, infected patients over the winter season of 2009 accounted for a staggering 16% of the total population.

Tackling the challenge of such global pandemics requires new technology for rapid clinical diagnosis. To answer this need,

RIKEN’s Diagnosis Technique Detects H1N1 at Record SpeedToshihisa Ishikawa and colleagues at the RIKEN OSC developed the RT-SmartAmp assay, a technique to rapidly detect the 2009 pdm influenza A(H1N1) virus from patient swab samples. By combining both reverse transcriptase (RT) and isothermal DNA amplification reactions in one step, the RT-SmartAmp assay does away with the need for RNA extraction and PCR reaction. The researchers adapted the RT-SmartAmp

technique using a fluorescent primer to specifically detect the 2009 pdm influenza A(H1N1) virus within 40 minutes, without cross-reacting with the seasonal A(H1N1), A(H3N2), or B-type (Victoria) viruses.

The effectiveness of the RT-SmartAmp method was confirmed in clinical research carried out at Japanese hospitals during the period of October 2009 to January 2010, where it outperformed standard diagnosis tests in both speed and sensitivity. Of a total 255 clinical samples, 140 (54.9%) were identified as 2009 pdm A(H1N1)-positive by RT-SmartAmp, compared to only 110 (43.1%) detected by standard diagnostic tests. In 72.8% of all 140 infection-positive cases, the RT-SmartAmp assay detected the presence of the pdm influenza virus within 24 hours of fever onset.

Taken together, these results set a new standard for infection diagnosis speed, providing a highly-effective tool for rapidly detecting sub-types of the H5N1 virus and oseltamivir-resistant influenza viruses and promising support in the battle to prevent global pandemic infection.

of even more complex systems, such as programmable molecular assembly lines and sophisticated sensors.

11

BIOBOARD

F or the past decade, scientists have been pursuing cancer treatments that exploit RNA interference — or RNAi, a

phenomenon that offers a way to shut off malfunctioning genes with short strands of RNA. However, delivering the RNA efficiently remains a big stumbling block to the field. Usually, short interfering RNA (siRNA) is quickly broken down inside the body by enzymes that defend against infection by RNA viruses.

“It’s been a real struggle to try to design a delivery system that allows us to administer siRNA, especially if you want to target it to a specific part of the body,” says Paula Hammond, the David H. Koch Professor in Engineering at MIT.

Hammond and her colleagues have now come up with a novel delivery vehicle in which RNA is packed into microspheres so dense that they withstand degradation until they reach their destinations. The new system, published on February 26th in the journal Nature Materials, knocks down expression of specific genes as effectively as existing delivery methods, but with a much

A large-scale study analyzing metabolic compounds in rice grains conducted by researchers at the RIKEN Plant

Science Center (PSC) and their collaborators has identified 131 rice metabolites and clarified the genetic and environmental factors that influence their production. The findings provide a natural way to bioengineer improved rice grain varieties by selectively increasing production of useful metabolites, boosting the nutritional value of crops.

As one of the most important staple crops, rice plays a central role in supplying the nutrients needed to keep the world population healthy. The nutritional value of rice crops is determined by the types and quantities of metabolites they contain, which are strongly affected by environmental and genetic factors. Understanding these factors is crucial to increasing nutritional value, but the complex relationship between genes and

New Hotspot on Rice Chromosome Could Yield Improved Varieties

plant metabolism makes this a formidable challenge.

At the heart of this challenge are so-called quantitative train loci (QTL), stretches of DNA which contain or link to the genes for a phenotypic trait, in this case metabolite levels. To breed lines of rice which produce more of a specific metabolite (for example one that boosts its nutritional value), you have to know which DNA regions are involved and in what role. This is hard because metabolite levels are controlled by many different QTLs and also strongly influenced by the environment.

To solve this problem, researchers at the PSC teamed up with their collaborators at the National Institute of Agrobiological Science (NIAS) to analyze rice grain metabolomic QTL (mQTL) using state-of-the-art mass spectroscopy pipelines developed at the PSC. Analysis of 85 experimental lines of rice

specially bred for QTL analysis, prepared by the NIAS researchers and harvested in 2005 and 2007, yielded a total of 759 metabolite signals. From these, the team identified 131 metabolites, including amino acids, lipids, and flavonoids, and identified a total of 801 mQTLs around the rice genome.

Most important of all, the team showed that while the levels of most metabolites they identified are influenced mainly by environmental factors, genetics can sometimes play a stronger role: coordinated control of amino acids was linked to an mQTL “hotspot” on chromosome 3, while variation of flavonoid levels was linked to genetic factors. Published in The Plant Journal, the findings promise a future of faster, more effective breeding techniques for rice, and mark a major step toward a healthier, better-fed world.

OTHER REGIONS

Delivering RNAi the Spongy Waysmaller dose of particles.

Such particles could offer a new way to treat not only cancer, but also any other chronic disease caused by a “misbehaving gene,” believes Hammond, who is also a member of MIT’s David H. Koch Institute for Integrative Cancer Research. RNA interference holds huge potential for a number of disorders besides cancer, including neurological and immune diseases.

To get the RNA to its destination, Hammond’s method packages the RNA as one long strand that would fold into a tiny, compact sphere. She used an RNA synthesis method known as rolling circle transcription to produce extremely long strands of RNA made up of a repeating sequence of 21 nucleotides. Those segments are separated by a shorter stretch that is recognized by the enzyme Dicer, which chops RNA wherever it encounters that sequence.

As the RNA strand is synthesized, it folds into sheets that then self-assemble into a very dense, sponge-like sphere. Up to half a million copies of the same RNA sequence can be packed into a sphere with a diameter

of just two microns. Once the spheres form, the researchers wrap them in a layer of positively charged polymer, which induces the spheres to pack even more tightly (down to a 200-nanometer diameter) and also helps them to enter cells.

After the spheres enter a cell, the Dicer enzyme chops the RNA at specific locations, releasing the precious cargo of siRNA sequences. These special sequences can encode instructions to shut off any gene of interest, making the method ideal for diseases that involve “misbehaving genes”. The really exciting part of the work according to Peixuan Guo, director of the NIH Nanomedicine Development Center at the University of Kentucky, is the development of a new self-assembly method for the RNA particles.

Hammond now plans to design microspheres coated with polymers that specifically target tumor cells or other diseased cells. Her team is also working on spheres that carry DNA, for potential use in gene therapy.[Source: MIT News Office]

12

EYE ON CHINA

In an effort to establish itself at the forefront of global biotechnology, China has recently announced the

National Program on Bioscience Technology Development 2010-2020. The program, which was issued by the Ministry of Science and Technology, reveals a series of ambitious, very specific goals for developing a world-class biotech industry in China. The stakes are high, and the goals lofty: China wants to produce three to five top biotech scientists with international reputations and Nobel Prize potential by 2020.

By 2020, the National Program plans to:

1. Produce 3-5 world-class scientists in bioscience, especially in the areas of biofuel and treatment of major diseases;

2. Build a “biotech talent pyramid,” focusing on five major areas, including development

China Unveils World-Class Biotech Ambitions of world-class scientific talent, first-rank innovation and teams, leadership, industry expertise, and biotech management;

3. Cultivate 30-50 world-class innovators a n d a n u m b e r o f i n n o v a t i o n teams in the areas of: genomics and functional genomics, stem cells and tissue engineering, transgenic plants and animals, cloned animals, and neuroscience;

4. Produce 300-500 leaders in the field, 30 ,000-50 ,000 spec ia l i s t s , and 300,000 bio-industry experts in the areas of bio-medicine, bio-agriculture, bio-manufacturing, and bio-environment; cultivate and develop 3000-5000 senior managers in biotechnology.

Approximately 250,000 people are already working in various biotechnology

fields in the PRC. Yet the field is stifled by a number of persistent issues:

1. A general shortage in the total number of biotechnology experts in China;

2. A lack of top-tier scientists;

3. A shortage in the number of people who return to China after they go abroad to study;

4. A lack of innovative research and innovative entrepreneurs.

The program more than doubles the number of people who will be working in biotechnology – from 250,000 to at least 550,000. To encourage innovation, China will have to provide financial support to its innovative companies, but perhaps even more important is the need to reward its top brains in the life sciences.

13

EYE ON CHINA EYE ON CHINA

T he National Foundation for Cancer Research (NFCR) recently announced that Dr. Zhen-Yi Wang and Dr. Zhu

Chen have been awarded the 7th Annual Szent-Györgyi Prize for Progress in Cancer Research for their innovative research that led to the successful development of a new therapeutic approach to acute promyelocytic leukemia (APL).

By combining traditional Chinese medicine with Western medicine, Drs. Wang and Chen have provided dramatic improvement in the five-year disease-free survival rate of APL patients - from approximately 25 percent to 95 percent - making this therapy a standard of care for APL treatment throughout the world, and turning one of the most fatal diseases into a highly curable one.

“I am so glad to see that the efforts we have devoted to research on leukemia these last several decades have led to solid clinical benefits to APL patients around the world,” said Dr. Wang. “This award will inspire us as we continue our efforts to find more effective therapies to treat cancers.”

“This is a great honor for Dr. Wang and me; it is quite humbling to know that our respected colleagues from many scientific disciplines have selected us for this prestigious award,” said Dr. Chen, who also serves as China’s Minister of Health. “Scientists across the globe are working every day to cure cancer. I hope our work may continue to inspire others.”

A clinical researcher at the Ruijin Hospital in Shanghai in the early 1980s, Dr. Zhen-Yi Wang performed the first successful therapy on APL patients using all-trans retinoic acid (ATRA), which significantly increased the survival rate of patients with APL. Dr. Zhu Chen, a former student of Wang, made major contributions to the identification of the molecular mechanisms of both ATRA and arsenic trioxide in APL. He also demonstrated in clinical trials that arsenic trioxide, a compound used as a traditional Chinese medicine for over 2,400 years, is effective against APL. Since the 1990s, Drs. Wang and Chen have worked together to conduct clinical trials combining ATRA and arsenic trioxide to treat APL

patients, with great success.“Drs. Wang and Chen have quite

literally changed the face of medicine for patients suffering from APL. Their combined work has saved countless lives and will continue to save many more lives around the world both today and in future generations,” said Dr. Beatrice Mintz, Fox Chase Cancer Center, Chair of the 7th Selection Committee of Szent-Györgyi Prize and winner of the 6th Annual Albert Szent-Gyorgyi Prize. “Terminal differentiation of cancer cells has been an important focus in my research, and I am very happy about the successful clinical application of this principle by Drs. Wang and Chen. I cannot imagine a better testament to the outcomes of investing in cancer research than what these two distinguished scientists have achieved.”

“In keeping with the spir it of nonconformity for which NFCR founder Albert Szent-Györgyi is known, the selection of Drs. Wang and Chen has a significant meaning for those who work in the trenches

of cancer research each day,” said Sujuan Ba, Ph.D., co-chair of the Szent-Györgyi Prize Selection Committee and chief operating officer of NFCR. “True scientific discovery comes from innovative ideas and dedicated research. These two scientists are inspirational, as they both have devoted their lives to this work that will impact the world for generations to come.”

The annual Szent-Györgyi Prize for Progress in Cancer Research was established by the National Foundation for Cancer Research to recognize outstanding scientific achievements in the war against cancer and to honor pioneering scientists who have made extraordinary contributions in cancer research. The focus of the Prize is on the critically important role that basic science plays in cancer research and in its application to cancer therapies. The Prize, which includes a $25,000 honorarium, will be presented to Dr. Wang and Dr. Chen at an award ceremony March 6, 2012 at The Westin Times Square in New York City.

Chinese Scientists Awarded Cancer Research Prize

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EYE ON CHINA

I n a new study published online in Nature Biotechnology, researchers from Beijing Genomics Institute (BGI),

the world’s largest genomics organization, reported evidence of extensive RNA editing in a human cell line by analysis of RNA-seq data, demonstrating the need for new robust methods to identify important post-transcriptional editing events.

RNA editing is a normal but not yet fully understood process in which small nucleotide changes occur after DNA has been transcribed into RNA. It is an integral step in generating diversity and plasticity of cellular RNA signature as a post-transciptional event that recodes hereditary information. RNA editing is an important area in the post-genomic era for its role in determining protein structure and function. It has become increasingly important in genetic research.

Last year, a study published in Science (Li, et al. Science, May 19, 2011) reported a large number of sequence differences between mRNA and DNA in the human transcriptome. This finding was startling because it implied that there might be a still undiscovered mechanism of ‘RNA editing’ that could disrupt the central dogma

BGI Researchers Uncover Extensive RNA Editing in Human Transcriptome

and affect our understanding of genetic variation. However, this view was strongly contested by other scientists because of the technical issue and lack of academic rigour, such as sequencing error or mis-mapping. In this latest study, BGI researchers developed a more rigorous pipeline for approaching these problems and answered some of the concerned questions, which contributed to paving way for the further studies of this field.

They obtained the whole-transcriptome data by RNA-seq from a lymphoblastoid cell line of a male Han Chinese individual (YH), whose genome sequence was previously reported as the first diploid genome of Han Chinese. RNA-seq, also known as “Whole Transcriptome Shotgun Sequencing”, is a recently developed approach on transcriptome profiling that uses deep-sequencing technologies with the advantages of high-throughput data, low background, high sensitivity and repeatability. In a paper published in 2009 in Nature Reviews Genetics (http://www.nature.com/nrg/journal/v10/n1/abs/nrg2484.html#top), RNA-seq is referred to as a revolutionary tool in transcriptomics.

“We used RNA-seq in the study to identify post-transcriptional editing events, and developed a computational and comprehensive pipeline to find the human RNA editing sites,” said Zhiyu Peng, the leading author of the paper and Vice Director of Research & Cooperation Division of BGI. The pipeline was used to identify the extensive RNA editing from genome and whole transcriptome data by screening RNA-DNA differences of the same individual through successive quality control filters.

Through this pipeline, BGI researchers identified 22,688 RNA editing events, and found most editing events (~93%) convert adenosine (A) into inosine (I), which in turn is read as guanosine (G), in consistence with known editing mechanisms based on adenosine deaminase acting on RNA (ADAR). They also found 44 editing events in microRNAs (miRNA), suggesting there is a potential connection between RNA editing and miRNA-mediated regulation. Researchers also found in the study evidence of other types of nucleotide changes, but these were validated at lower rates.

“These findings demonstrate this multifilter molecular pipeline is an excellent approach in this study,” said Peng. “With the multiple filters, false positive results can be controlled or eliminated while identifying RNA editing events, providing a more accurate and effective method to extensively analyze RNA editing. We now plan to apply this new methodology to larger-scale deep sequencing studies for more comprehensive analysis and profiling of editome, including studies with additional physiologically relevant samples.”

“The evidence of extensive RNA editing identified in a human transcriptome underscores the necessity of an effective method to fully detect these events in order to further advance our understanding of human development and normal pathophysiological condition,” said Jun Wang, Executive Director of BGI. “With continual improvement of the new approach, we believe this could be achieved in the near future.”

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J ohns Hopkins recently launched the China STEM (Science, Technology, Engineering and Medicine) program

to apply the Chinese language to growing academic fields for career purposes in partnership with Nanjing University. Beginning in the summer of 2012, this eight-week long summer program of study will take place in the Hopkins-Nanjing University Center for Chinese and American Studies.

The China STEM program will offer undergrads, graduate students, professionals, post-doctoral fellows and researchers courses that seek to bridge the language barriers between Chinese and American scientists and engineers. It will present these students with the language skills necessary to excel in a career in science, technology, engineering or medicine. The program offers

Johns Hopkins Commences New China STEM Program

a mixture of intensive language training in small classes, research seminars, personal interactions with Chinese scholars and scientists and experiential visits to other institutions such as laboratories, hospitals, and academic institutions in Nanjing and Beijing.

The China STEM program does not align with a specific major, but seeks to appeal to students interested in medical, public health or engineering careers. Scholars and professionals in these technical fields have continuously lacked proficiency in the Chinese language despite the growing necessity for such a capability. Given China’s growing economic and political clout on the world-stage, skills in the Chinese language are likely to become more important to

specialists in medicine, public health and engineering. Currently, there are many classes in “Business Chinese”, but these may be inadequate in equipping students with skills to communicate effectively with their Chinese counterparts on matters of science and technology. The China STEM program fills the gap in such language training.

Johns Hopkins aims to train a new generation of bilingual American scientists who would have a distinctive edge in a globalized scientific landscape, one in which China is set to play a dominant role. The China STEM program is a tacit recognition of China’s impending dominance in science and technology, and all indicators point to more universities in Europe and the USA setting up similar exchange programmes.

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EYE ON CHINA

T he ‘Father of hybrid rice,’ Professor Yuan Longping, was conferred the 2011 Mahathir Science Award recently

at the Kuala Lumpur Convention Center, Malaysia. The Award, given out annually by the Mahathir Science Award Foundation (MSAF), was presented by former prime minister Tun Dr. Mahathir Mohamad and witnessed by the Minister of Science, Technology and Innovation, Dr. Maximus Johnity Ongkili; Chairman of MSAF, Tun Ahmad Sarji Abdul Hamid; and President of the Academy of Sciences Malaysia, Tan Sri Dr. Ahmad Tajuddin Ali FASc .

Yuan won the award in recognition of his contributions to rice breeding, resulting in the innovative development of hybrid rice, a staple food of the tropics that has revolutionized global rice production and sustainability.

China’s disastrous famine from 1958 to 1961 was a turning point in Yuan’s life and led him to concentrate his research on the development of high-yielding rice. Since then, Yuan devoted himself to the research and development of a better rice breed. In

Father of Hybrid Rice Conferred 2011 Mahathir Science Award

1964, he happened to find a natural hybrid rice plant that had obvious advantages over others. Greatly encouraged, he began to study the elements of this particular type.

The biggest problem by then, was having no known method to reproduce hybrid rice in mass quantities, and that was what Yuan set out to solve. In 1964, Yuan created his theory of using the probably-existing naturally-mutated male-sterile rice individuals for the creation of reproductive hybrid rice species, and in two years he managed to find a few individuals of such male-sterile rice that he predicted could be used for his research. Subsequent experiments proved his theory feasible, which was his most important contribution on hybrid rice.

More problems followed but Yuan persevered. The first experimental hybrid rice species cultivated didn’t show any significant advantage over common ones, so he suggested to crossbreed rice with its further relative: the wild rice. In 1970, he found an important species of wild rice that he needed for the creation of high-yield

hybrid rice species. In 1973, in cooperation with others, he was finally able to establish a complete process of creating and reproducing high-yield hybrid rice species.

The next year they successfully cultivated a type of hybrid rice species which had great advantages. It yielded 20 percent more per unit than that of common ones, making China a worldwide leader in rice production. For this achievement, he was dubbed the “Father of Hybrid Rice.”

At present, as many as 50 percent of China’s total rice fields grow Yuan Longping’s hybrid rice species and yield 60 percent of the rice production in China. Due to Yuan’s hard work, China’s total rice output rose from 5.69 billion tons in 1950 to 19.47 billion tons last year, about 300 billion kilograms more have been produced over the last twenty years. The annual yield increase is enough to feed 60 million people.

The “Super Rice” Yuan is now working on and has yields 30 percent higher than those of common rice. A record yield of 17,055 kilograms per hectare was registered in Yongsheng County in Yunnan Province in 1999.

A ffymetrix, Inc. (NASDAQ:AFFX) recently announced that its GeneChip® System 3000Dx v.2 (GCS

3000Dx v.2) has been approved by China’s State Food and Drug Administration (SFDA) for in vitro diagnostic use. The GCS 3000Dx v.2 is the first microarray instrument system to be granted SFDA registration for array-based diagnostics for enabling personalized medicine. China has more than 2,000 clinical centers that will now have access to the only SFDA-cleared microarray platform for clinical testing.

The molecular diagnostic market in China is the fastest growing in the world and represents a significant growth opportunity for Affymetrix in Asia. “We are delighted to be the first SFDA-cleared microarray platform, as this will enable us to expand into the clinical diagnostics applications,”

Personalized Medicine Receives Boost in Chinasays Chris Barbazette, Vice President, Commercial Operations International Markets at Affymetrix.

The GCS 3000Dx v.2 microarray platform has a proven record of successful development and commercialization through partnership via the Powered by Affymetrix™ (PbA) program. A number of companies are developing molecular diagnostic tests in cancer, cardiovascular diseases, and inherited disorders based on the Affymetrix GeneChip platform. More than ten tests are in the pipeline for regulatory clearance. Two FDA-cleared tests (Roche AmpliChip® CYP450 Test and Pathwork® Diagnostics’ Tissue of Origin Test) and three CE-IVD marked tests, including Skyline Diagnostic’s AML test, are currently on the market. These tests and Affymetrix’ own solutions for cytogenetics, cancer, and pharmacogenomics are part of an

increasing menu of clinical applications that can be run on the SFDA-cleared GeneChip System.

“Having an SFDA-cleared system and a wide-range of clinical tests will enable physicians in China to bring personalized medicine to their patients faster,” says Dr. Ming Zhang at Hangzhou Bio-San Biochemical Technologies Company.

“This registration clearance is a significant accomplishment for Affymetrix and supports our global clinical strategy. It connects us more closely to physicians in China wanting to utilize clinically relevant genomic biomarkers that improve their patients’ health and wellness,” said Andy Last, Executive Vice President of Genetic Analysis and Clinical Applications Business Unit at Affymetrix.

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G eneNews Limited (TSX:GEN), a molecular diagnostics company focused on developing blood-based

biomarker tests for the early detection of diseases and personalized health management, today announced that it has signed a memorandum of understanding (“MOU”) with Shanghai Biochip Co. Ltd. (“SBC”), a leading Chinese engineering centre for advanced microarray and gene expression profiling technologies, to enter into a strategic alliance to establish the first Sentinel Centre for Personalized Medicine in China.

GeneNews and SBC would jointly establish and manage the Sentinel Centre for Personalized Medicine to co-develop and commercialize additional products based on GeneNews’ proprietary platform technology, the Sentinel Principle®. In addition, the MOU sets forth the main criteria to be incorporated into definitive development, distribution and license terms under which SBC would obtain non-exclusive rights to

GeneNews & Shanghai Biochip to Establish First Sentinel Centre for Personalized Medicine

market and sell GeneNews’ ColonSentry™ test in China. A formal agreement will be negotiated and executed before the end of the Company’s third fiscal quarter. Financial terms were not disclosed.

“We are focused on bringing leading edge technologies to the people of China to improve healthcare outcomes,” stated Dr. Yuchen Chen, Director, Business Development of SBC. “We are looking forward to working with GeneNews to establish the world’s first Sentinel Centre for Personalized Medicine to advance the development and commercialization of innovative, non-invasive tests targeted at early disease detection in China.”

“Shanghai Biochip’s expertise in gene expression profiling platforms and well-established clinical network provide a strong foundation for us to jointly establish a state-of-the-art research and development core capability in China that is aligned with our pipeline development and commercialization

objectives. We look forward to building a successful strategic alliance with Shanghai Biochip,” said Gailina J. Liew, President & Chief Operating Officer of GeneNews.

The Sentinel Principle®, a platform technology discovered and developed by GeneNews, is based on the concept that all clinical conditions and body states, including those resulting from disease or in response to treatment, generate characteristic gene expression signatures in the blood as a result of the constant and dynamic physiological interaction of blood with the cells, tissues and organs of the human body. This technology is the basis of GeneNews’ initial product, ColonSentry™, the world’s first blood test for colorectal cancer, and the SentinelGx™ suite of services. GeneNews’ broad global patent portfolio includes issued foundational patents and pending patents in diverse disease areas such as cancer, cardiovascular, neurological and inflammatory conditions.

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EYE ON CHINA

D r. GAN Mingzhe from Suzhou Institute of Nano-Tech and Nano- Bionics, Chinese Academy of

Sciences (SINANO, CAS) invented a novel scalable multi-channel microfluidic chip for bacterial suspension culture. In this device, culture media in 32 parallel and identical culture chamber loops are driven by a series of micropumps formed with a single set of control channels. The culture media indifferent culture loops are demonstrated to cycle at the same speed, which ensures identical growth environments in all culture loops. Suspended cultivations of microbial strains such as E.coli, Bacillus subtilis, Pseudomonas stutzeri and Zymomonas mobilis are carried out to demonstrate general applicability of the chip for microbial culture. The simple chip design and low-cost fabrication allow for an efficient microbial suspension culture. This work has been published on Lab on a Chip (http://pubs.

SINANO Invents New Microfluidic Chip for Bacterial Culture

rsc.org/en/Content/ArticleLanding/2011/LC/c1lc20670b).

Microbes are widely used in chemical, food, pharmaceutical, and health care industries as well as new industries including environmental remediation, green chemistry, sustainable manufacturing, biomass energy and resources, and low-carbon biodiesel industries. Improving the microbial production strains offers great opportunity to gain more profits with less capital outlay, thus realizing sustainable development of these industries. Shake flasks and tubes account for over 90% of microbe culture experiments. However, current flask culture techniques have poor screening efficiency due to the large numbers of flasks, bulky shaking beds, large fluid volume, intensive human labor and expensive equipment.

Based on their work, Dr. GAN and his colleagues developed a second-generation microbial culture chip (Figure 2A). A

high degree of integration of 120 culture chambers (50 nL each) in a 7.5 cm× 5 cm chip is achieved. A faster circulating flow rate enables the suspension culture of various microbial strains including bacteria as well as yeast (Figure 2B). Simpler fabrication process allows for future low-cost industrial chip production. This work has been published on Small. (http://onlinelibrary.wiley.com/doi/10.1002/smll.201102322/abstract)

Both works are parts of the project “Microbial Screening and Analysis System”. The project is to develop an integrated microfluidic system for microbial screening and optimization, which could greatly speed up strain selection process in microbial industry. Currently, the research is focusing on developing rapid micro-analysis technology for microbial metabolites. The work is supported by the Hundred Talents Program of CAS and the Knowledge Innovation Program of CAS.

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APBN: What is the current focus of your research? How is it important?

John Yu: I have assembled an interdisciplinary team of investigators in Academia Sinica, Taiwan, and formed a cohesive, interrelated team work to gather a better understanding of stem cell biology. Currently, our research aims to develop innovative technology platforms for identification of unique markers and regulators for normal and cancer stem cells. We employ glycomics/glycoproteomics, phosphoproteomics, coupled with scFv antibody library, and high throughput shRNA screening to interrogate differential profiles of glycans/glycoproteins in stem cells and differentiated cells and their downstream regulatory control. Now, launching the newly found markers of human embryonic stem cells (ESCs) or breast cancer stem/initiating cells (CSCs), we are trying to (a) investigate the role of these markers and their downstream regulators in the maintenance of the self-renewal capability and differentiation potentials of hESC, lung stem cells, and their CSCs, (b) isolate normal lung stem cells and identify CSCs for lung adenocarcinoma, (c) study the inter-relationship of biomarkers between normal stem cells and cancers, and (d) target CSCs, especially of breast cancer, and to explore the possibility of development of cancer therapeutics targeting CSCs with these biomarkers via specific antibodies or siRNA.

APBN: What are its potential applications?

John Yu: Specific surface markers are valuable for monitoring the culture and behavior of stem cells and their status of differentiation. Many commonly used stem cell markers were directly adopted from phenotypic characteristics of other cell types with limited insight into the specific stem cells under study. Therefore, it is important to identify new and more markers of stem cells. On one hand, this current project should help to sort out from the newly found surface markers those molecules that will facilitate the isolation and expansion of stem cells, and also help to shed light on the functional roles of these molecules in stem cells and CSCs. On the other hand, the innovative technology platforms developed in this project will not only promote the understanding of the relationship between stem cells and cancers, but also foster the development of cancer detection and therapeutics targeting new glycans /glycoproteins.


W ith his wealth of knowledge and formidable experience, biotech scientist, Dr John Yu decided to move back to his homeland with aspirations to widen Taiwan’s biotechnology horizons and bring his country to prominence on the international stage of biotech research.

Dr Yu gave up his position as Director of Experimental Hematology, Department of Molecular and Experimental Medicine at The Scripps Research Institute to move back to his native Taiwan, determined to bring to his country international standards, experience and knowledge. He is currently the Distinguished Research Fellow at Institute of Cellular and Organismic Biology, and Genomics Research Center. In an exclusive interview, APBN’s Sonal finds out what makes this biotech scientist tick.

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APBN: What are the milestones achieved by your lab?

John Yu: Based on MALDI-MS and MS/MS analyses, we have found a close association of the expressions of glycosphingolipids (GSLs) on the surface of hESCs with differentiation. It is believed that the newly found glycans present in hESCs could be candidates for cancer detection and glycan-targeted therapy of human tumors. SSEA-3 is a pentasaccharide (2Galβ1→ 3GalNAcβ1→ 3Gala1→ 4Galβ1→ 4Glcβ1) and serves as the precursor of Globo H. SSEA-4 is the sialylated derivative of SSEA-3 (NeuNAca2 -SSEA-3). On the other hand, Globo H, a known biomarker for cancers, was highly expressed uniquely in hESCs and its antibodies reacted with both Globo H and SSEA-3. Therefore, these markers for hESCs are perhaps the targets of therapy for cancers. We also showed that there was a striking switch in the core structures for globo- and lacto-series GSLs during differentiation of hESCs into embryoid body, neural progenitors, or endodermic cells, suggesting a close association of GSLs in hESCs with lineage-specific differentiation (PNAS 107 22564 2010; Stem Cells, 29: 1995 2011).

More recently, we employed glycoproteomics and glycan analysis to analyze the glycoprotein/glycan expression patterns for hESCs. It was found that seven newly found surface glycoproteins were also highly expressed in breast cancer stem/initiating cells. For example, ESC02 which is uniquely present in hESCs is highly expressed (at least 7 folds) in the paired cancer stem cell subpopulation of breast cancer from different patients versus non-CSC. When transduced with shESC02, the expression level of Sox2 in shESC02 cells was reduced to 40%, whereas no significant changes were seen in Oct4 and Nanog expression. In addition, gene knockdown of ESC02 expression leads to decrease in the self-renewal of both hESCs and mammary spheres of primary breast cancer from patients. Such ESC02 silencing in ESCs also results in developmental skewing toward endoderm/mesoderm differentiation in vitro and in vivo. Therefore, these results warrant the development of ESC02 as therapeutic agents for cancer, because knockdown of its expression fulfills the major requirements for new therapeutic agents of cancer: i.e. cell arrest and differentiation induction. Furthermore, shedding of ESC02 occurs to the medium after incubation with breast cancer cells. Thus, we had developed a platform to study the inter-relationship between stem cells and cancer and to develop new cancer detection method and glycan-targeted therapy.

APBN: Academia Sinica collaborates a lot with foreign universities. How is it helpful?

John Yu: Academia Sinica, the most preeminent academic institute in Taiwan was founded to pursue research excellence. There are 24 institutes and 7 research centers in Academia Sinica under three divisions: Mathematics and Physical Sciences, Life Sciences, as well Humanities and Social Sciences. At present, there are more than 1,500 researchers with Ph.D. degree and an annual budget over US$400 million. Among 246 academicians, there are six Nobel Laureates. The purpose of academic research is to improve human life. Academia Sinica will continue its tradition of pursuing solid research while exploring new knowledge and will remain focused on the needs of society in the hope of enriching human civilization.

Academia Sinica, with its own state of arts and expertise, also participates in the pursuit of research excellence worldwide. For example, President Chi-Huey Wong will visit the United States in late March 2012 along with 12 scholars from Academia Sinica. He will attend the “Future of Glycoscience” report committee meeting organized by the U.S. National Academy of Sciences; attend the Academia Sinica – UC Berkeley Symposium; give a lecture and sign a Collaborative Agreement at the University of California at San Diego; and receive Arthur C. Cope Award from the American Chemical Society. Such a visit is expected to boost academic cooperation between Taiwan and the United States.

In 2011, Academia Sinica President Emeritus Yuan-Tseh Lee officially took up the reigns of the International Council for Science (ICSU). He was inaugurated as President of this international organization during the General Assembly held in Rome, which was attended by over 300 ICSU representatives from around the world. Another example is the Yuan-Tseh Lee Array for Microwave Background Anisotropy (AMiBA) that is a forefront instrument for

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research in cosmology. This project is led, designed, constructed, and operated by extensive international and domestic scientific and technical collaborations. The facility is sited on Mauna Loa in Hawaii, USA, at an elevation of 3,400m to take advantage of higher atmospheric transparency and minimum radio frequency interference.

APBN: You were the Director of Experimental Hematology, Department of Molecular & Experimental Medicine, The Scripps Research Institute from 1998 to 2002 before you decided to move to Taiwan. What made you return to your country after working in US for more than three decades?

John Yu: In 2002, Professor Yuan-Tseh Lee, then President of Academia Sinica, offered me the position of the Director for Institute of Cellular & Organismic Biology (2002-2009) and the chief of Stem Cell Program. It was a great opportunity for me to do something for my country. In addition to pursue my own research, I set up a Stem Cell Program at the Academia Sinica, established the Taiwan Society for Stem Cell Research (www.tsscr.org.tw) and became its founding President.

I was also elected to serve in ISSCR International hESC Guidelines Task Force, Government Affairs Committee, and the Steering Committee of Stem Cell Network in Asia-Pacific regions. I had been in USA for more than 30 years; but the contributions which lead to personal satisfaction in your own motherland have been far greater than what one can expect to receive in the States.

APBN: Please elaborate on your role with the government of Taiwan with regard to Research and Development of Science in Taiwan

John Yu: In Taiwan, I serve on many advisory boards concerning biomedical research and biotechnology development.

I returned to Taiwan in 2002 and began efforts to establish a new Stem Cell Program in Academia Sinica. After successfully setting up a dynamic multidisciplinary program at the Genomics Research Center with approximately 80 personnel by May 2003, a newly renovated research lab was opened, providing the most up-to-date facilities for basic and translational research. The major emphasis of this Program is to pursue cutting edge stem cell research such as specific markers, pluripotency, and epigenetic control of normal and cancer stem cells. The members of this team participate in several national stem cell flagship programs from other institutes/universities.

One of my pioneer research achievements is the identification of a rare subpopulation of lung cells with the characteristics of pulmonary stem cells and the demonstration of their susceptibility to SARS-CoV and avian flu H5N1 (PNAS 103: 9530-9535, 2006). I am also trying to develop innovative technology platforms for identification of unique surface markers, and investigate the control of genetic regulators and epigenetic profiling affecting stem cells under the Flagship Program by combining several strategies including glycoproteomics and glycan analysis, phosphopeptide enrichment/LC/MS/MS analysis.

In 2005, Taiwan Society for Stem Cell Research was established and I became the founding President for this society (2005-2010). I was also the chief architect for the National Strategic Plan for Stem Cell Research in Taiwan since 2006. I was charged with the responsibility to set up National Strategy Plan for stem cell research. Under my leadership, a 4-point strategic plan was established: 1) establishing national flagship programs, 2) implementation of ethics and regulation, 3) setting-up infrastructures and resources, and 4) development of international collaborations. This strategy plan lays out the basic foundation and infrastructural support for stem cell research in Taiwan.

APBN: Taiwan is essentially a Buddhist country. Did the religious and social values affect Stem Cells research in anyway.

John Yu: The general public and patients in Taiwan hold enormous anticipation towards the development and success of stem-cell research. Since stem cell technology touches upon

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important ethical issues and public policy that required resolution, we had actively engaged with the public and hold extensive educational activities for the community. For example, we organized hearings for general public and proactively participated in the meetings organized by churches or Buddhist monks and nuns, to discuss ethical issues of stem cell research and potential values for the society. At the government level, we also formed a special Committee for Biotech, Ethics, & Social Communication 2010 to open up societal discourse and inform government policy related to new developments of biomedical technologies, such as stem cell research. Furthermore, since I was a part of ISSCR International hESC Guidelines Task Force, I actively advised the Department of Health in Taiwan to modify guidelines for stem cell research which now adheres to the international policy and regulations worldwide.

In order to ensure a role of Taiwanese stem cell researchers in international collaboration, Department of Health Executive Yuan in Taiwan had announced on August 9, 2007 a new regulatory guideline, “Policy Instructions on the Ethics of Human Embryo and Embryonic Stem Cell Research”. In addition, I actively worked to get National Legislation of Human Embryo and Embryonic Stem Cell Research Act approved, which was eventually passed in Executive Yuan on July 24, 2008. People in Taiwan have now realized that the development of ethical standards and practices is a critical catalyst for international collaboration and for research to be accepted ethically and for it to be validated by the scientific community.

John Yu, M.D., Ph.D. is Distinguished Research Fellow at Institute of Cellular and Organismic Biology, and Genomics Research Center; and the Chief of Stem Cell Program, AS, Taiwan. Dr. Yu was the Director for Institute of Cellular and Organismic Biology (2002-2009). He is the founding President for Taiwan Society for Stem Cell Research (www.tsscr.org.tw). Dr. Yu has been elected to serve in ISSCR International hESC Guidelines Task Force, Government Affairs Committee, the Steering Committee of Stem Cell Network in Asia-Pacific regions, and visiting Professor for Stem Cell Biology, Kumamoto University, Japan. He was Director of Experimental Hematology (1998-2002) at Scripps Research Institute, USA. He received an Established Investigatorship Award from American Heart Association, etc.

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Its Associated Risks and Promise to Mankind

What are stem cells and why they have been in vogue?

S tem cells are the precursors of all cells in the human body. They have the ability to replicate themselves and

to repair and replace other tissues in the human body. Stem cells are, therefore, very special, powerful cells not only in humans but also in animals. Besides humans and animals, plant stem cells have also been known since a long time. Plant stem cells are innately undifferentiated cells located in the meristems of plant capable of giving rise to a steady supply of precursor cells to form differentiated tissues and organs in the plant. However, the details of human and animal stem cells will be discussed. Moreover, the translational aspect of stem cells is precisely known as regenerative medicine [1]. In other words, medicine that involves growing of new cells, tissues and organs to replace or repair those damaged by injury, disease or aging is called regenerative medicine.

What makes stem cells special is that they are regenerative and malleable. The malleability of the stem cells which enables them to give rise to any other cell type in human body is technically known as plasticity of stem cells [2, 3]. And this is the key property of stem cells due to which the stem cells have been in vogue.

Bipasha Bose

Nanyang Technological University, Singapore

What are different sources and types of stem cells? Just as there are many different types of specialized or differentiated cells in the body, there are many different types of stem cells in the body. Hierarchically, stem cells are classified as totipotent, pluripotent and multipotent [4, 5,]. This hierarchy is decided on the plasticity of stem cells. The most plastic type of stem cells which have the capability to theoretically give rise to

any possible cell type including the entire organism are called totipotent, for example-zygote and oocyte. Totipotent stem cells are followed by a lesser plastic type of stem cells known as pluripotent stem cells. Pluripotent stem cells are capable of giving rise to any possible cell type but not the entire organism. The examples of pluripotent cell types are embryonic stem cells and induced pluripotent stem cells. Followed by pluripotent stem cells are a much least plastic version of stem cells which can give rise to only a limited number of cell types preferably from the same germ layer from which the stem cell

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itself is derived. Such least plastic version of stem cells is called multipotent stem cells. Examples of multipotent stem cells are precisely adult stem cells like mesenchymal cells and stromal cells.

In the end, where the plasticity of stem cells end, we have the non-plastic version of cells known as unipotent cell types [4]. Such cells are not stem cells, but are rather somatic cells representing majority of cell types in the human body. Somatic cell types are most of the times capable of dividing and giving rise to only its own cell type. A typical example is glial cell found in nervous system, a muscle cell etc.

So, broadly speaking the basic types of stem cells are embryonic and adult/ somatic stem cells. Also, their sources are so called from embryonic origin (mostly totipotent and pluripotent stem cells) and somatic tissue (adult stem cells like-hematopoietic stem cells, muscle stem cells, liver stem cells etc.) respectively.

History of stem cell researchIn the early 1900’s European researchers realized that the various type of blood cells e.g white blood cells, red blood cells and platelets all came from a particular ‘stem cell’. However, it was not until 1963 that the first quantitative descriptions of the self-renewing activities of transplanted

mouse bone marrow stem cells were documented by Canadian researchers, Ernest A McCulloch and James E Til [6]. Then in 1998 James Thomson, a scientist at the University of Wisconsin in Madison, successfully removed cells from spare embryos at fertility clinics and grew them in the laboratory. He launched stem cell research into the limelight, establishing the world’s first human embryonic stem cell line which still exists today [7]. Much prior to the establishment of human embryonic stem cells (hESC), mouse embryonic stem cells (mESC) were generated from mouse embryos by Evans and Kaufman (1981) and Martin (1981) thereby, establishing a platform for pluripotent stem cells [8, 9]. Then the breakthrough discovery came when artificial counterparts of natural pluripotent stem cells were generated in a dish by Shinya Yamanaka and his group from Japan in 2007[10]. Yamanaka’s group demonstrated for the first time that by introducing four pluripotency genes named Oct4, Sox2, Klf4 and c-Myc, somatic cells can be made to behave like pluripotent stem cells. The stem cells generated by this method were termed as Induced Pluripotent stem cells (iPSC). Then onwards, the standard technology for generating iPSC have been used globally to generate pluripotent stem cells from various cell types and used in the basic sciences research as well as applied aspect of stem cell biology.

History of stem cells in translational medicineStem cell therapy named bone marrow transplantation have been done since past five decades or so to treat life threatening diseases like leukemia and blood related disorders. Georges Mathé, a French oncologist, performed the first bone marrow transplant in 1959 on five Yugoslavian nuclear workers whose own marrow had been damaged by irradiation caused by a Criticality accident at the Vin a Nuclear Institute, but all of these transplants were rejected. Stem cell transplantation was pioneered using bone-marrow-derived stem cells by a team at the Fred Hutchinson Cancer Research Center from the 1950s through the 1970s led by E. Donnall Thomas, whose work was later recognized with a Nobel Prize in Physiology or Medicine in 1990 [11]. The first physician to perform a successful human bone marrow transplant on a disease other than cancer was Robert A. Good at the University of Minnesota in 1968 [12]. Then onwards since a decade ago, transplantation of cord blood stem cells obtained from umbilical cord also have been used for transplantation purpose [13].

Moreover, rest of the stem cell therapies especially the ones derived from embryonic stem cells and induced pluripotent stem cells have safety issues like likelihood of teratomas formation in addition to ethical concerns for human embryonic stem cells. Overcoming all that issues, a California based company Geron Corporation on January 23, 2009, got the US Food and Drug Administration clearance for the initiation of the first clinical trial of an embryonic stem cell-based therapy on humans. The trial aimed to evaluate the drug GRNOPC1, embryonic stem cell-derived oligodendrocyte progenitor cells, on patients with acute spinal cord injury [14]. The trial was discontinued in November 2011 so that the company could focus on therapies in the “current environment of capital scarcity and uncertain economic conditions”.

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Risk factors associated with stem cell therapyAlthough stem cell therapy holds the promise to treat degenerative diseases, cancer and repair of damaged tissues for which there are currently no or limited therapeutic options. Despite the great promise, there are still many questions regarding the safe application of stem cell therapy. The risk of stem cell therapy is categorized based on both theoretical concerns and examples of adverse observations. Risks associated with stem cell therapy depend on many risk factors. A risk is defined as a combination of the probability of occurrence of harm and the severity of that harm [15, 16]. Examples of risk factors are the type of stem cells used, their procurement and culturing history, the level of manipulation and site of injection. Because of the variety of risk factors, the risks associated with different stem cell based medicinal products may differ widely as well. For an adequate benefit/ risk assessment of a stem cell based medicinal product, all important identified risks (i.e. risks or adverse events identified in clinical experience) as well as potential/ theoretical risks (e.g. non-clinical safety concerns that have not been observed in clinical experience) should be thoroughly evaluated at the start and during the development of a stem cell based therapy. Specific examples are- Embryonic stem cells (ESC) are pluripotent cells that have the ability to differentiate into derivatives of all three germ layers (endoderm, mesoderm,

and ectoderm). Theoretically speaking, ESCs demonstrate complete differentiation ability to a particular cell type to offer therapy. But, under most of the practical situations, there remain chances of undifferentiated ESC population. This undifferentiated ESC and iPSC population poses a risk of forming teratomas (tumors) when transplanted for cell therapy purpose [17]. Likewise, this particular risk of tumor formation of stem cell is attributed to the resemblance of stem cells to that of cancer cells, such as long life span, relative apoptosis resistance and ability to replicate for extended periods of time. Therefore, stem cells may be considered potential candidates for malignant transformation [18, 19, 20, 21]. In addition, similar growth regulators and control mechanisms are involved in both cancer and stem cell maintenance. This is probably why tumor formation is often seen as a key obstacle to the safe use of stem-cell based medicinal products. Site of administration is another potential risk factor wherein mouse ESC transplanted into a homologous species caused highly malignant teratocarcinomas at the site of administration, while xenotransplantation in rats resulted in migration and differentiation of the mESC [22]. Such observation was also recorded in case of human ESC. Another risk is in vitro expansion and culture of stem cells can change the characteristics of the stem cell due to intracellular and extracellular influences [23]. Every cell division has a small chance of introducing deleterious mutations and mechanisms to correct these alterations may not function as adequate (e.g. cell cycle arrest, DNA repair), or at all (e.g. immune recognition)

occur during in vitro culture. Cell culture induced copy number changes and loss of heterozygosity have been reported for human ESC lines. In principle, such changes may cause transformation of a cell into a tumorigenic phenotype and may contribute to increased tumor formation. This risk due to in vitro expansion also holds true for non embryonic/adult stem cells.

Will stem cells be able to deliver breakthrough contribution to alleviate human suffering and disease in near or far future?Although a number of stem cell therapies exist, but most are at experimental stages, risky or costly. Medical researchers anticipate that adult and embryonic stem cells will soon be able to treat cancer, Type 1 diabetes mellitus, Parkinson’s disease, Huntington’s disease, Celiac Disease, cardiac failure, muscle damage and neurological disorders, and many others [24].

Neverthe less , before s tem ce l l therapeutics can be applied in the clinical setting, more research is necessary to understand stem cell behavior upon transplantation as well as the mechanisms of stem cell interaction with the diseased/injured microenvironment.

References1. Mason C, Dunnill P (2008). “A brief definition of regenerative medicine”. Regenerative Medicine 3 (1): 1–5

2. Krause, Diane S.; Theise, Neil D.; Collector, Michael I.; Henegariu, Octavian; Hwang, Sonya; Gardner, Rebekah; Neutzel, Sara; Sharkis, Saul J. (2001). “Multi-Organ, Multi-Lineage Engraftment by a Single Bone Marrow-Derived Stem Cell”. Cell 105 (3): 369–77

3. Mountford, JC (2008). “Human embryonic stem cells: origins, characteristics and potential for regenerative therapy.”. Transfus Med 18: 1–12

4. Hans R. Schöler (2007). “The Potential of Stem Cells: An Inventory”. In Nikolaus Knoepffler, Dagmar Schipanski, and Stefan Lorenz Sorgner. Human biotechnology as Social Challenge. Ashgate Publishing, Ltd. p. 28. ISBN 9780754657552

5. Mitalipov S, Wolf D (2009). “Totipotency, pluripotency and nuclear reprogramming.”. Adv Biochem Eng Biotechnol 114: 185–99

6. Becker, A.J.; McCulloch, E.A.; Till, J.E. (1963). “Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells”. Nature 197 (4866): 452–4

7. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic Stem Cell Lines Derived from Human Blastocysts”, Science, 282(5391):1145-7

8. Evans M, Kaufman M (1981). “Establishment in culture of pluripotent cells from mouse embryos”. Nature 292 (5819): 154–6

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Dr. Bipasha Bose is currently a Post Doctoral Research Fellow in School of Biological Sciences, Nanyang Technological University, Singapore. She did her Master’s in Biotechnology in 1997 from G.B.Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, India. Thereafter, Dr. Bose obtained her PhD in 2004 from Advanced Centre for Treatment, Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai, India. Her PhD work was on Applied Biology focusing primarily on signaling pathways involved during malignant transformation of cancer cells from normal healthy cells induced by non-permitted food adulterant used in third world countries. Then onwards Dr. Bose moved into the field of stem cell biology primarily due to the similarity in cancer cells and stem cells. She first worked in industry based stem cell research in Regenerative Medicine unit in the company named Reliance Life Sciences Pvt. Ltd, Navi Mumbai, India. While working in this company, Dr. Bose co-worked with scientists for pioneering the derivation of human embryonic stem cells in Indian subcontinent from human blastocyst obtained from in vitro fertility clinics. Moreover, she also led the diabetes sub-group for deriving beta islet cells from human embryonic stem cells in the same company. After four years of translational research in stem cells, Dr. Bose moved back to academia to work in Stem Cell Institute as a post-doctoral fellow in Catholic University Leuven, Belgium under the mentorship of Prof. Catherine Verfaillie. Her post-doctoral work in Belgium involved differentiation of human embryonic cells into liver and kidney cells and also basic research on pluripotency genes in ES cells. Furthermore, in her current post-doctoral work she is investigating various aspects of aging and muscle derived stem cells, culture induced pluripotency of muscle stem cells, occurrence of muscle like cells in the liver. She owns six peer reviewed publications and one international patent to her credit.

About the Author

9. Martin G (1981). “Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells”. Proc Natl Acad Sci USA 78 (12): 7634–8

10. Takahashi K, Yamanaka S (2006). “Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors”. Cell 126 (4): 663–76

11. Thomas ED, Lochte HL Jr, Lu WC, Ferrebee JW (1957). Intravenous infusion of bone marrow in patients receiving radiation and chemotherapy. N Engl J Med 257:491-496

12. Good RA (2002).Cellular immunology in a historical perspective. Immunol Rev 85:136-58

13. Laughlin MJ, Barker J, Bambach B, et al. (2001). Hematopoietic engraftment and survival in adult recipients of umbilical-cord blood from unrelated donors. N Engl J Medicine 344:1815-22

14. Keirstead HS, Nistor G, Bernal G, et al. (2005). “Human embryonic stem cell-derived oligodendrocyte progenitor cell transplants remyelinate and restore locomotion after spinal cord injury”. J. Neurosci. 25 (19): 4694–705

15. ISO 14971:2007. [http://www.iso.org/iso/iso_catalogue.htm]

16. Guideline on risk management systems for medical products for human use, EMEA/CHMP/96268/2005. [http://www.ema.europa.eu]

17. Knoepfler PS: Deconstructing stem cell tumorigenicity: A roadmap to safe regenerative medicine (2009). Stem Cells 27:1050-1056

18. Shih CC, Forman SJ, Chu P, Slovak M (2007). Human embryonic stem cells are prone to generate primitive, undifferentiated tumors in engrafted human fetal tissues in severe combined immunodeficient mice. Stem Cells and Development 16:893-902

19. Amariglio N, Hirshberg A, Scheithauer BW, Cohen Y, Loewenthal R,Trakhtenbrot L, Paz N, Koren-Michowitz M, Waldman D, Leider-Trejo L, et al.(2009). Donor-derived brain tumor following neural stem cell transplantation in an ataxia telangiectasia patient. PLoS Medicine 6:0221-0231

20. Werbowetski-Ogilvie TE, Bosse M, Stewart M, Schnerch A, Ramos-Mejia V,Rouleau A, Wynder T, Smith MJ, Dingwall S, Carter T, et al(2009). Characterization of human embryonic stem cells with features of neoplastic progression.Nature Biotechnology 27:91-97

21. Strauss R, Hamerlik P, Lieber A, Bartek J (2012). Regulation of Stem Cell Plasticity: Mechanisms and Relevance to Tissue Biology and Cancer. Mol Therapy. 2012 Feb 7. doi: 10.1038/mt.2012.2. [Epub ahead of print]

22. Erdo F, Buhrle C, Blunk J, Hoehn M, Xia Y, Fleischmann B, Focking M,Kustermann E, Kolossov E, Hescheler J, et al.(2003). Host-dependent tumorigenesis of embryonic stem cell transplantation in experimental stroke. Journal of Cerebral Blood Flow and Metabolism, 23:780-785

23. Narva E, Autio R, Rahkonen N, Kong L, Harrison N, Kitsberg D, Borghese L,Itskovitz-Eldor J, Rasool O, Dvorak P, et al.(2010). High-resolution DNA analysis of human embryonic stem cell lines reveals culture-induced copy number changes and loss of heterozygosity. Nature Biotechnology 28:371-377

24. Power C and Rasko JEJ (2011). Promises and Challenges of Stem Cell Research for Regenerative Medicine. Ann Intern Med 155:706-713

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Introduction

A bility to reprogram differentiated somatic cells by transferring genetic material dates back to 1957 with

first experiments performed in frogs1. However, full implications of reprogramming technology were only demonstrated with Dolly, the sheep four decades later2. These and other early studies demonstrated that reprogramming via nuclear transfer or incorporation of genetic materials was feasible in higher mammalian systems. In 2006, Takahashi and Yamanaka for the first time demonstrated that this reprogramming technology could be utilized to turn back the developmental clock of adult somatic cells to their pluripotent/embryonic state by utilizing 4 important nuclear transcriptional factors3. These reprogrammed cells or induced pluripotent stem cells (iPSCs) demonstrated features similar to embryonic stem cells (ESC) that were derived from inner cell mass of an embryo.

While the advent of human embryonic stem cell (hESC) technology had fostered great optimism, their pluripotency is however not easily controlled. Only if harnessed appropriately, could they provide powerful models of human development, disease manifestation and regenerative materials for cell replacement therapies. This vision has been partially realized with the provision of disease-specific hESC lines from afflicted embryos identified in pre-implantation diagnostic screening programs4, but it is

*Please send requests and correspondence to: Dr Ashish MehtaNational Heart Centre Singapore,17, Third Hospital Avenue,Mistri Wing, Singapore 168752Email: [email protected]: +65 65365357Fax: +65 62263972


Ashish Mehta* and Winston ShimResearch and Development Unit, National Heart Centre Singapore

Figure 1. Phase contrast photomicrograph shown a typical human induced pluripotent stem colony grown in feeder free system.

limited because some diseases cannot be diagnosed during pre-implantation diagnosis5. However, this limited applicability of hESC in disease modeling and to some extend in cell therapy (ethical and immunological issues) was overcome with the advent of hiPSCs. One of the most promising applications for hiPSC derived cell types, apart from regenerative therapy is disease modeling. Today there are various emerging reports that show generation of patient specific iPSC for disease modeling including cardiomyopathies6-11.

Generation of diseased human iPSC for cardiomyopathiesHopes of establishing disease-specific pluripotent stem cells as a starting material for generating surrogate models for human diseases are understandably high. This in principle means that any disease could be modeled “in a dish” through iPSC technology. While several methods exist to reprogram somatic cells like viral (retrovirus or lentivirus)

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or non-viral (episomal, minicircles, proteins and RNA); viral vectors are associated with major drawback of genomic integration, insertional mutations, residual transgene expression and tumorigenesis12, making viral free systems a better choice for various applications including cell therapy. Our group has previously demonstrated that skin cells could be effectively reprogrammed using viral free systems to generate iPSCs (Figure 1) that efficiently differentiate into functional cardiomyocytes12.

It is also important to know that most disease phenotypes are only observed in lineage-committed or differentiated cells derived from iPSCs, but not in the iPSCs. Thus, pertinent information on the pathogenesis of a disease may only be obtained from hiPSCs that have been differentiated in vitro to disease-relevant cell types such as cardiomyocytes for cardiac diseases. There are a number of cardiomyopathies reported clinically. In theory, it is possible to generate iPSCs with same disease phenotypes in vitro, it is however not practical currently for a number of reasons. One of the main points to consider is the complexity of disease manifestations. While some disease may be easy to study, others may not be conducive or not adequately manifested in vitro. Monogenic diseases are an attractive target because there is a clear

relation between genotype and phenotype of the disease. In case of cardiomyopathies, arrhythmogenic or heart rhythm disease in which a single point mutation in genes results in a defective function could be easily evaluated for modeling. In general, most common arrhythmogenic diseases are due to mutations in the ion channel genes, an integral part of cardiac physiology, which could be evaluated by electrophysiology techniques. A number of pioneering proof

DISEASE METHOD OF REPROGRAMMING GENE MUTATION REFERENCE

LQTS 1 Retrovirus KCNQ1 gene, R190Q mutation 6

LQTS 2 Retrovirus KCNH2 gene, A614V mutation 7

LQTS 2 Lentivirus KCNH2 gene, A561T mutation 8

LQTS2 Retrovirus KCNH2 gene, R176W mutation 15

LQTS 2 non-viral KCNH2 gene, A561V mutation 16 (Unpublished)

CPVT1 Retrovirus RYR2 gene, 7447T>A mutation 17

CPVT 2 Retrovirus RYR2 gene, S406L mutation 18

Timothy Syndrome Retrovirus CACNA1C gene, G406R mutation 19

Studies with patient iPSC derived cardiomyocytes for cardiac disease modeling

Abbreviations: LQTS: Long QT syndrome, CVPT: Catecholaminergic polymorphic ventricular tachycardia

of principle studies by our group and others have shown iPSC derived cardiomyocytes from patients with inherited arrhythmogenic diseases, most notably different subtypes of long QT syndrome, recapitulate the clinical phenotype in the culture dish6, 8, 13. Table 1 summaries the studies till date performed on cardiovascular disease utilizing hiPSC technology. These publications focusing on arrhythmic syndromes convincingly confirm our expectations that disease modeling will

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References1. Gurdon JB, Elsdale TR, Fischberg M. Sexually mature individuals of Xenopus laevis from the transplantation of single somatic nuclei. Nature 1958;182(4627):64-5.

2. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH. Viable offspring derived from fetal and adult mammalian cells. Nature 1997;385(6619):810-3.

3. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007;131(5):861-72.

4. Pickering SJ, Minger SL, Patel M, Taylor H, Black C, Burns CJ, Ekonomou A, Braude PR. Generation of a human embryonic stem cell line encoding the cystic fibrosis mutation deltaF508, using preimplantation genetic diagnosis. Reprod Biomed Online 2005;10(3):390-7.

5. Colman A. Induced pluripotent stem cells and human disease. Cell Stem Cell 2008;3(3):236-7.

6. Moretti A, Bellin M, Welling A, Jung CB, Lam JT, Bott-Flugel L, Dorn T, Goedel A, Hohnke C, Hofmann F, Seyfarth M, Sinnecker D, Schomig A, Laugwitz KL. Patient-specific induced pluripotent stem-cell models for long-QT syndrome. N Engl J Med 2010;363(15):1397-409.

7. Itzhaki I, Maizels L, Huber I, Zwi-Dantsis L, Caspi O, Winterstern A, Feldman O, Gepstein A, Arbel G, Hammerman H, Boulos M, Gepstein L. Modelling the long QT syndrome with induced pluripotent stem cells. Nature 2011;471(7337):225-9.

8. Matsa E, Rajamohan D, Dick E, Young L, Mellor I, Staniforth A, Denning C. Drug evaluation in cardiomyocytes derived from human induced pluripotent stem cells carrying a long QT syndrome type 2 mutation. Eur Heart J 2011.

be one of the main contributions of hiPSC technology to human health.

While modeling monogenic diseases is currently feasible; most cardiovascular diseases are complex or their clinical manifestations cannot be simply linked to any single genetic abnormality. For instance, ischemic or diabetic cardiomyopathies that are secondary to multifactorial conditions may be difficult to model. Furthermore, their evaluation may not be as straightforward as in case of arrhythmogenic diseases. A particular challenge would also be to evaluate diseases that have a “late onset” in humans, this is mainly because hiPSC derived cardiomyocytes are more fetal-like in phenotype. Though in-principle the technology has immense potential as a tool to study and model various cardiomyopathies, associated factors like generation of pure populations of cardiomyocytes, maturity of cardiomyocytes may become significant hurdles in understanding complex and late onset cardiovascular diseases.

Personalized drug regimes Besides revealing mechanistic insights of the modeled disease, modeling using iPS technology opens an avenue to study new chemical entities in treating the disease. Transgenic models in rodents have advanced our basic understanding of human diseases,

but they often not reproduce clinical pathophysiology appropriately. For example, humans exhibit significant differences in cardiac electrophysiology when compared to mice. These differences can be easily observed in their cardiac action potential or ECGs. Mice have a much shorter action potential than humans. The QT interval in mouse is between 50-100 ms, but 400 ms in human14. These variations are mainly due to differences in handling of potassium currents in both the species14. Thus, patient derived hiPSCs provide additional dimension to drug screening for various diseases.

Despite their current limitations m e n t i o n e d a b o v e , i P S - d e r i v e d cardiomyocytes appear well-suited for drug evaluations as they respond appropriately to various pharmaceutical agents in a manner similar to adult cardiac cells. This property of iPSC derived cardiomyocytes could be exploited extensively to evaluate drugs for better health care. In fact, most of the current research papers on disease modeling have focused on drug evaluations. This shows that besides modeling of the disease, designing better drug molecules or testing efficacy of drug molecules that previously could never be tried on patients could now be successfully performed. This may be very important clinically in the management of cardiac patients, especially those suffering with heart rhythm problems like LQTS, where selection of drug regimes is an important

feature in managing the disease. It should be noted that in order to develop a new drug, the efficacy of the drugs should be tested on large population of patients, which would be highly challenging and expensive. In contrast, iPSC technology could be used on individual patient making personalized drug regimens possible. The field of drug testing and disease modeling are still in its infancy and a lot of further work is required to establish these concepts into clinical scenarios.

Concluding remarks Disease modeling with patient-specific hiPSCs will generate a wealth of information and data that when combined with genetic analysis may allow early and more accurate predictions and diagnosis of disease and its progression. Moreover, disease modeling is likely to provide an understanding of why different individuals show variations in response to drugs, which could significantly contribute to disease management and pharmaceutical industry. Lastly, this research will likely ignite a paradigm shift among patients, doctors and society at large and these changes may bring unforeseen challenges to existing governing policies in regard to privacy and ethics involving human materials and data generated. It is imperative to be mindful of such implications as iPSC technology is poised to usher in a new paradigm in clinical practice.

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9. Israel MA, Yuan SH, Bardy C, Reyna SM, Mu Y, Herrera C, Hefferan MP, Van Gorp S, Nazor KL, Boscolo FS, Carson CT, Laurent LC, Marsala M, Gage FH, Remes AM, Koo EH, Goldstein LS. Probing sporadic and familial Alzheimer’s disease using induced pluripotent stem cells. Nature 2012;482(7384):216-20.

10. Pasca SP, Portmann T, Voineagu I, Yazawa M, Shcheglovitov A, Pasca AM, Cord B, Palmer TD, Chikahisa S, Nishino S, Bernstein JA, Hallmayer J, Geschwind DH, Dolmetsch RE. Using iPSC-derived neurons to uncover cellular phenotypes associated with Timothy syndrome. Nat Med 2011;17(12):1657-62.

11. Song B, Sun G, Herszfeld D, Sylvain A, Campanale NV, Hirst CE, Caine S, Parkington HC, Tonta MA, Coleman HA, Short M, Ricardo SD, Reubinoff B, Bernard CC. Neural differentiation of patient specific iPS cells as a novel approach to study the pathophysiology of multiple sclerosis. Stem Cell Res 2012;8(2):259-73.

12. Mehta A, Chung YY, Ng A, Iskandar F, Atan S, Wei H, Dusting G, Sun W, Wong P, Shim W. Pharmacological response of human cardiomyocytes derived from virus-free induced pluripotent stem cells. Cardiovasc Res 2011;91(4):577-86.

13. Malan D, Friedrichs S, Fleischmann BK, Sasse P. Cardiomyocytes obtained from induced pluripotent stem cells with long-QT syndrome 3 recapitulate typical disease-specific features in vitro. Circ Res 2011;109(8):841-7.

14. Davis RP, van den Berg CW, Casini S, Braam SR, Mummery CL. Pluripotent stem cell models of cardiac disease and their implication for drug discovery and development. Trends Mol Med 2011;17(9):475-84.

15. Lahti AL, Kujala VJ, Chapman H, Koivisto AP, Pekkanen-Mattila M, Kerkela E, Hyttinen J, Kontula K, Swan H, Conklin BR, Yamanaka S, Silvennoinen O, Aalto-Setala K. Model for long QT syndrome type 2 using human iPS cells demonstrates arrhythmogenic characteristics in cell culture. Dis Model Mech 2011.

16. Mehta A, Lester GL, Sudibyo Y, Wong P, Liew R, Shim W. Derivation and Characterization of Transgene Free Induced Pluripotent Stem Cell Derived Cardiomyocytes from Asian Patient With Long QT Syndrome. Unpublished 2012.

17. Fatima A, Xu G, Shao K, Papadopoulos S, Lehmann M, Arnaiz-Cot JJ, Rosa AO, Nguemo F, Matzkies M, Dittmann S, Stone SL, Linke M, Zechner U, Beyer V, Hennies HC, Rosenkranz S, Klauke B, Parwani AS, Haverkamp W, Pfitzer G, Farr M, Cleemann L, Morad M, Milting H, Hescheler J, Saric T. In vitro modeling of ryanodine receptor 2 dysfunction using human induced pluripotent stem cells. Cell Physiol Biochem 2011;28(4):579-92.

18. Jung CB, Moretti A, Mederos YSM, Iop L, Storch U, Bellin M, Dorn T, Ruppenthal S, Pfeiffer S, Goedel A, Dirschinger RJ, Seyfarth M, Lam JT, Sinnecker D, Gudermann T, Lipp P, Laugwitz KL. Dantrolene rescues arrhythmogenic RYR2 defect in a patient-specific stem cell model of catecholaminergic polymorphic ventricular tachycardia. EMBO Mol Med 2011.

19. Yazawa M, Hsueh B, Jia X, Pasca AM, Bernstein JA, Hallmayer J, Dolmetsch RE. Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. Nature 2011;471(7337):230-4

Dr. Ashish Mehta obtained his Ph.D (2003) from Defence R and D Establishment, Ministry of Defence, India in Biochemistry with sub-specialization in biochemical toxicology. Following his post-doctoral work in Leishmanial apoptosis, Dr Mehta joined a private company as Senior Research Scientist (2006) for developing protocol involving human ES cells and their differentiated cell types for drug screening. He is currently a Senior Research Scientist at National Heart Centre, Singapore focusing on generation of normal and disease specific induced pluripotent stem cells using non-viral methods for cardiac disease modeling and possible applicability for drug screening. Dr Mehta has published more than 30 publications in international peer reviewed journal and is also attached with a number of journals as a reviewer.

Dr Winston Shim obtained his Ph.D. (2001) from National University of Singapore in molecular biology and then joined National Heart Centre, Singapore in the same year. His research interest is in adult bone marrow mesenchymal stem cells in cardiac repair. He was among the first in the world to demonstrate ex vivo differentiation of adult stem cells to cardiomyocytes. He is currently Scientific Director, Research and Development Unit, National Heart Centre Singapore; Principal Investigator, Stem Cell Laboratory, National Heart Centre Singapore; Assistant Professor, DUKE-NUS Medical Graduate School, Singapore. His current area of research is to develop better models to understand and treat cardiac patients, using adult as well as induced pluripotent stem cells derived cardiomyocytes. Dr Shim has extensive publications in cardiac regenerative and gene therapy.

About the Authors

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T he idea behind stem cell therapy is the application of cells to restore damaged tissues in the body and

thereby restoring the health of the patient who has lost significant use of their bodily functions. Some of the types of diseases being addressed include restoring the eye sight, repairing bone and cartilage damage, re-establishing heart function after a heart attack, reinstating lost insulin production and reconnecting neural functions in various debilitating diseases such as spinal cord injury, Parkinson’s and stroke. Scientific progress is telling us that stem cells may provide some of these special features if they can be guided to the specialised cell types, but to cross over from the lab dish to the patient treatment, these specialised cells will have to be produced on a massive scale. Another application is to use stem cells to screen for molecules that may facilitate differentiation to the tissue of interest or to screen out drugs that may have harmful side effects.

To achieve this, one of the enabling technologies is bioprocessing which is the development of the knowhow and capabilities to grow billions to trillions of cells consistently for animal studies, clinical trials and eventually therapy in patients.

Microcarrier cultures of stem cellsThe Bioprocessing Technology Institute has developed simple, yet rigorous and

Bioprocessing for Stem Cell Therapy From the Lab into the Log Phase

Steve OhBioprocessing Technology Institute, Agency for Science Technology and Research (A*STAR)

a)

b)

Figure 1. a) Clusters of pluripotent stem cells growing as large aggregates surrounding several rod shaped microcarriers Scale bar 100 microns. b) Multipotent stem cells growing as monolayers on spherical shaped microcarriers.

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increased the yield and was generally applicable for differentiating both hESC and hiPSC cell lines to cardiomyocytes. These cardiomyocytes express cardiac muscle, show normal electrophysiology and are now being implanted into animal models of cardiac infarct to see if they can restore heart function. The cells can also be applied for drug screening of potentially cardiotoxic compounds.

Secondly, we have taken the expanded MSC and placed them in scaffolds as well as fibrin glue mixed with hydroxyapatite and implanted them into mouse models. Surprisingly, it was shown that MSC produced from microcarrier cultures were able to produce much more calcium per

reproducible methods for growing vast populations of stem cells such as human embryonic stem cells (hESC) obtained from an embryo, human induced pluripotent stem cells (hiPSC) obtained by genetic modification of mature differentiated cells, and multipotent stem cells (MSC) usually obtained from bone marrow. Furthermore, these cells have been directed to become functional cell types of the body such as beating clusters of heart cells, nerve cells and bone cells in scaffolds. BTI approached the problem by growing the cells on the surface of tiny structures called microcarriers (ranging from 10 to 200 micron in diameter or cylinders of about 100 micron in length), and are about 10 times larger than the size of the stem cells to which they can attach to. These microcarriers are positively charged and coated with extracellular matrix proteins (like laminin and vitronectin) such that the cells can rapidly anchor to the surface, spread and grow (Fig.1). The microcarrier platform which operates in stirred reactors allows increase of cell amounts simply by increase of volume, as compared to the conventional method which scales up linearly by increasing the number of plastic vessels. In these new culture conditions, it is important to demonstrate that the cells still retain the stem cell characteristics such as the expression of protein markers by the cells (mAb 84, Tra-1-60, and Oct4 for hESC and hiPSC and Stro-1, CD 73, CD 90 and CD 105 for MSC) similar to that obtained in classical laboratory conditions.

Furthermore, we can increase the cell density from a typical yield of 1 million cells/ml on tissue culture flasks, to 6 million cells/ml in microcarrier cultures. In a 1 litre bioreactor culture, it is possible to generate 6 billion hESC or hiPSC in serum free media, which is equivalent to manually handling 80 tissue culture flasks of 75mls. Thus so far, we see no upper limit to the highest density that can be achieved, but we are only limited by the culture media supply to the cells. For MSC, we have achieved 0.8 million cells/ml in microcarrier cultures compared to 0.2 million cells/ml in traditional tissue culture flasks, using serum supplemented media.

Stem cell differentiation in bioreactorsThe next stage after cell expansion is to differentiate them towards the specific cell type of interest. In three different examples, we have demonstrated that microcarriers can enable an increased efficiency of differentiation. After screening for a variety of microcarriers, we determined that 10 micron microcarriers were the best in generating cardiomyocytes yielding 0.6 cardiomyocytes / hESC. Subsequently, we developed a serum free media with nutritional supplements which further

Figure 2. Spinner flasks for suspension microcarrier of stem cells and their differentiated progenitors

Fig. 3. a) Differentiated cardiomyocytes stained with alpha-actinin. b) Neurons stained with b-tubulin III. Scale bars are 10 micron. c) Micro-CT of ectopic bone formed by MSC.

a) b)

c)

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cell and generate 60% higher bone volume than MSC produced from plastic tissue culture flasks. Thus, we have developed a simplified method of producing large numbers of MSC in a bioreactor that have a better characteristic of bone formation for potential therapeutic application.

Thirdly, we developed a process in spinner flask cultures (Fig. 2) for serial expansion and differentiation of both hESC and hiPSC to neuroprogenitors, which are the precursors of nerve cells. During the neural differentiation phase, we achieved a massive 6 to 11 fold increase in the yield of neuroprogenitors compared to the tissue

Improvements in yields of production of stem cells and their differentiated progenitors.

HUMAN STEM CELLS

DEFINED MEDIA

2D CULTURES(MILLION CELLS/ML)

MICROCARRIER CULTURES

(MILLION CELLS/ML)

FOLD IMPROVEMENT

hESC Yes 1 to 2 2 to 6 2 to 3 fold

hiPSC Yes 0.8 to 1.2 3 to 6 3 to 5 fold

hMSCNo, with serum

0.1 to 0.2 0.6 to 0.8 3 to 4 fold

DIFFERENTIATED CELLS

DEFINED MEDIA

2D OR EB CULTURES(CELLS/HPSC)

MICROCARRIER CULTURES

(CELLS/HPSC)

FOLD IMPROVEMENT

Cardiomyocytes Yes 0.1 to 0.2 0.3 to 0.6 3 to 6 fold

Neuroprogenitors Yes 32 to 53 333 to 371 6 to 11 fold

Ectopic bone Not

applicable19 (bone volume) 12 (bone volume) 1.6 fold

culture method. Final cell densities exceeded 1 billion cells/ml compared to 1 to 2 million cells/ml in tissue culture and 80% of them expressed the neuroprogenitor surface marker PSA-NCAM. The neuroprogenitors were further differentiated to functional neurons and can potentially be used for studying the restoration of lost functions in Parkinson’s disease, as well as for screening for compounds that differentiate these cells to neurons, or astro-glial lineages.

Table 1 summarises the yield of microcarrier processes compared to the 2D culture equivalent methods for producing anchorage pluripotent or multipotent

stem cells as well as their differentiated progenitors. Factors of improvement range from 2 to 11 fold depending on the cell type expanded and tissue formed.

ConclusionThus, we believe that microcarrier cultures are very versatile for stem cell expansion and differentiation; with a potential for consistently generating huge quantities of anchorage dependent stem cells and their progenies for applications in drug screening and future cell therapies in a cost effective manner.

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Dr Steve Oh received his PhD in Biochemical Engineering in Birmingham University, UK in 1990 and he is currently Principal Scientist and Associate Director at the Bioprocessing Technology Institute (BTI), one of the biomedical research institutes under the Agency for Science Technology and Research (A*STAR). Since 2001, together with Dr. Andre Choo they have nucleated and grown the Stem Cell Group in BTI with particular focus on bioprocessing issues related to human embryonic stem cells (hESC) resulting in >50 published papers and 9 filed patents. The group’s main areas of research are stem cell characterization, serum free, feeder free culture and scale up. Most recently, the group has developed a microcarrier platform technology on defined extracellular matrices which is amenable for bioprocess control and potential commercial scale production of pluripotent hESC, hiPSC, human mesenchymal stem cells, cardiomyocytes, neural stem cells and osteoblasts. His vision is to create viable bioprocesses for cell therapy, and drug development applications with allogeneic stem cells.

About the Author

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Stem Cell Group 2, BTI. Feb. 2012

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Sources of Neural Stem Cells for Clinical Application

C ell replacement therapy could aid i n a l l ev ia t ing symptoms o r even reversing progression of

neurodegenerative diseases. Over the last

Neural Stem Cells and Cell-based Approaches in Neurodegeneration

and Peripheral Nerve InjuriesYi-Chao Hsu and Ing-Ming Chiu

Division of Regenerative Medicine, Institute of Cellular and System Medicine, National Health Research Institutes, Miaoli, Taiwan

decade, convincing evidence has emerged of the capability of various stem cell populations to induce regeneration in animal models of Parkinson’s disease (PD), Huntington’s disease (HD) and Alzheimer’s disease (AD) along with multiple sclerosis and cerebral ischemia [Gogel et al., 2011]. Neural stem cells (NSCs) hold tremendous potential for neurodegenerative diseases.

This approach is not limited to the use of NSCs for transplantation, but includes the stimulation of endogenous stem cells and the multiple bioactive molecules that they express during reciprocal interactions with the diseased central and peripheral nerve systems. Several critical problems for NSC clinical application remain to be resolved: (i) the source of NSCs should be personalized;

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(ii) the isolation methods and protocols of human NSCs should be standardized; (iii) the clinical efficacy of NSC transplants must be evaluated in more adequate animal models and (iv) the mechanism of intrinsic brain repair needs to be better characterized. In addition, the ideal imaging technique for tracking NSCs would be safe and yield high temporal and spatial resolution, good sensitivity and specificity.

The ideal sources of NSCs for clinical application should better be personalized, tolerant to immune rejection, and resistant to tumorigenesis, easy to obtain and to amplify, effective in cellular or molecular replacement. The potential cell sources for central nervous system transplantation include fetal or adult NSCs [Tamaki et al., 2002; Uchida et al., 2000], embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs). NSCs can be expanded over a long period of time and as they are already neuralized (for example, committed to a central nervous system cell fate) there is no need for recapitulating early developmental signals that lead to neuroectodermal commitment. However, the transplantation of fetal NSCs into the adult brain is accompanied by numerous ethical, scientific and legislative hurdles [Mathews et al., 2008]. In addition, the prolonged culturing of NSCs leads to an ever increasing glial differentiation pattern at the expense of neuronal differentiation, which significantly reduces the therapeutic potential of fetal NSCs [Anderson et al., 2007]. Human ESCs can be manipulated to generate defined neuronal and glial lineages, thereby offering a major opportunity to study neurodevelopment and model neurological disease in vitro, as well as potentially having direct therapeutic applications in the field of regenerative neurology. However, certain challenges remain to be resolved before the promise of ESCs for neurological diseases can be fully realized, including the need to optimize survival, fate and function of neural derivatives upon both neural conversion and long-term differentiation in vitro and in vivo. Current protocols for the derivation of NSCs from ESCs require defined conditions; however, these conditions lead to significant cell death, in which the production of reactive oxygen species has a central role. Thus, neuralization protocols often contain antioxidants (which may increase the

propensity to accumulate genetic mutations), involve co-culture with stromal feeder layers or use B27 and/or N2 supplements. It is becoming increasingly clear that the more traditional stem cell systems using oxygen levels approximating room air (20%) are far from optimal, particularly with regard to neural specification and differentiation. Thus, recent protocol has been designed to generate NSCs and their regionally specified derivatives from ESCs using a physiological oxygen level of 3% (normoxia) [Stacpoole et al., 2011].

Flow cytometry and fluorescence-activated cell sorting (FACS) have been used successfully to resolve the complexity of lineage progress in the hematopoietic and other systems. Recently, FACS has been applied extensively in NSC biology, such as isolation of different precursor and progenitor populations from the central nervous system and peripheral nervous system. NSCs that had been isolated from brain tissues by cell-surface markers, such as CD133, or GFP expression driven by NSC-specific promoters. These promoters include Sox1, Sox2, Nestin and FGF1 [Hsu et al., 2009b; Lee et al., 2009]. NSCs thus isolated were cultured in the presence of growth factors and examined to determine whether they could expand to form neurospheres. The capacity to form neurospheres was defined as self-renewal. The potential for neural differentiation of these isolated cells upon withdrawal of growth factors or administration of inducing factors was used to determine multipotency.

Introduction of NeurodegenerationThe prevalence of neurodegenerative d i s e a s e s i s i n c r e a s i n g r a p i d l y . Neurodegenerative diseases such as AD, HD and PD trigger neuronal cell death through endogenous suicide pathways. Although progressive neuronal loss is a hallmark of neurodegenerative disorders, some neurological impairment may reflect dysfunction rather than loss of neurons. Abnormal protein assemblies seem to trigger vicious cycles of aberrant neuronal activity and compensatory alterations in neurotransmitter receptors and related signaling pathways that lead to synaptic deficits, disintegration of neural networks, and, ultimately, failure of neurological functions.

Recently, the U.S. President Obama administration plans to spend an additional $156 million over the next two years to help find an effective treatment for AD that affects more than five million Americans. The spending increase is intended to help in achieving a U.S. target set last month to find a way to treat or prevent AD by 2025. Current drugs help manage symptoms but so far no therapy can stop the progression of AD, which can start with vague memory loss and confusion before progressing to complete disability and death. Experts predict that without an effective treatment, the number of Americans with AD will double by 2050 and related healthcare costs could soar to more than $1 trillion a year.

Neurodegenerative diseases can disrupt molecular pathways, synapses, neuronal subpopulations and local circuits in specific brain regions, as well as higher-order neural networks. Abnormal network activities may result in a vicious cycle, further impairing the integrity and functions of neurons and synapses, for example, through aberrant excitation or inhibition. Neurodegenerative disorders such as AD and amyotrophic lateral sclerosis (ALS) are associated with microvascular dysfunction and/or degeneration in the brain, neurovascular disintegration, defective blood-brain barrier function and/or vascular factors. Microvascular deficits diminish cerebral blood flow and, consequently, the brain’s supply of

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oxygen, energy substrates and nutrients. Moreover, such deficits impair the clearance of neurotoxic molecules, non-neuronal cells and neurons. Recent evidence suggests that vascular dysfunction leads to neuronal dysfunction and neurodegeneration, and that it might contribute to the development of proteinaceous brain and cerebrovascular ‘storage’ disorders. Such disorders include cerebral β-amyloidosis and cerebral amyloid angiopathy, which are caused by accumulation of the peptide β-amyloid in the brain and the vessel wall, respectively, and are features of AD [Zlokovic, 2011].

PD is a degenerative disorder of the central nervous system. It was first described in 1817 by James Parkinson. Researchers believe that at least 500,000 people in the United States currently have PD, although some estimates are even higher. Society pays an enormous price for PD. The total cost to the nation is estimated to exceed $6 billion annually. The risk of PD increases with age, so analysts expect the financial and public health impact of this disease to increase as the population gets older. PD is characterized by an extensive loss of dopaminergic neurons in the substantia nigra, pars compacta, and their terminals in the striatum [Kish et al., 1988]. Although the etiology of idiopathic PD is not known, several predisposing factors for the dopaminergic depletion associated with the disease have been suggested, including programmed cell death, viral infection, and environmental toxins. As an effective treatment for PD, patients have been given L-dihydroxyphenylalanine (L-DOPA), a precursor of dopamine, but long-term administration of L-DOPA consequently produces grave side effects.

Introduction of Peripheral Nerve InjuriesThe incidence of peripheral nerve injury (PNI) in developed countries is estimated between 13 and 23 per 100,000 persons per year. Injuries to peripheral nerves result in partial or total loss of motor, sensory and autonomic functions in the involved segments of the body. Reinnervation of denervated targets can be achieved by

regeneration of injured axons or by collateral branching of undamaged axons in the vicinity. Nevertheless, these mechanisms do not provide for satisfactory functional recovery, especially after severe injuries. Peripheral nerve problems are common and encompass a large spectrum of traumatic injuries, diseases, tumors and iatrogenic lesions. The incidence of traumatic injuries is estimated as >500,000 new patients annually in the world. PNI results in loss of neural control in denervated segments of the body, and severe disabilities for the patients. Nerve regeneration usually does not allow for adequate target reinnervation and functional restitution. Neuronal response and axonal regeneration imply a complex interaction of cell types and changes in the expression of many molecules. Many experimental models have been used to gain knowledge on nerve regeneration and to develop strategies to promote recovery.

The failure of axons to regenerate following PNI results from decreased intrinsic properties of the neurons [Kadoya et al., 2009], the absence of neurotrophic factors or the presence of inhibitory factors in the environment. After PNI, axons and myelin sheaths distal to the lesion are degraded. The degenerative products are eliminated by the cooperative action of denervated Schwann cells and infiltrating macrophages. Wallerian degeneration serves to create a microenvironment favoring axonal regrowth. Schwann cells within the endoneurial tubes of the distal nerve dedifferentiate towards a non-myelinating proliferative phenotype that over-express growth factors, cell adhesion molecules and extracellular matrices. The axotomized neurons shift from a ‘transmitter’ state to a ‘regenerative’ state, so their axons generate growth cones that progress from the proximal stump into the distal nerve. Axonal regeneration requires an adequate substrate of trophic factors, provided by reactive Schwann cells, macrophages and the extracellular matrix within the degenerated nerve. The regenerative process, however, does not usually reconstitute a normal nerve structure neither allows for normal distal reconnection after severe lesions. Neuronal response and axonal regeneration require a complex interaction of several cell types and

changes in the expression of many molecules with variable spatial and temporal patterns. Therefore, a wide variety of methods are used in experimental studies, depending on the specific goals of each study [Navarro, 2009].

Current Therapeutic Applications of NSCs in Neurodegeneration and Peripheral Nerve InjuriesIt is widely anticipated that transplantation of stem/progenitor cells will provide effective therapies for many neurological diseases and injuries [Trounson et al., 2011]. Numerous encouraging animal studies have shown that stem or progenitor cell treatments can rescue some degree of neurological function after injury. Current clinical trials of human NSCs and ESCs are shown in Table 1. We further summarize the preclinical studies evaluating NSC therapy for peripheral nerve repair in Table 2. Only few studies have demonstrated direct evidence of cell replacement in injury or disease models that clearly explains the benefits observed after cell therapy. Many positive outcomes after cell therapy appear to be attributed to rescue of pre-existing tissue rather than repair or cell replacement per se. The paracrine action of growth factors, cytokines, and hormones that are secreted or released by transplanted cells has been shown to provide most of the benefits after stem/progenitor cell administration. This can be seen as a problem, since for many years we missed paracrine activity as a principal mechanism in cell therapy and the paracrine mechanism may be complicated. Alternatively, the situation can be viewed as an exciting opportunity to the new cell replacement therapy. Notably, a research group in Taipei Veteran General Hospital developed a novel spinal cord injury repair strategy. Using peripheral nerve grafts and FGF1 improves hindlimb locomotor function in spinal cord-transected rats [Cheng et al., 1996; Lee et al., 2002; Tsai et al., 2008]. Repaired spinal cords induce the expression of the M2 macrophage marker arginase I six

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to 14 days after repair and recruited large numbers of M2 macrophages to the graft area 10 days after repair [Kuo et al., 2007]. They further demonstrate that FGF1 induces IL-4 expression and that nerve grafts induce NGF and BDNF expression in transected spinal cords. A full repair strategy utilizes the beneficial effects of both FGF1 and nerve grafts simultaneously [Kuo et al., 2011].

The human FGF1 gene was first cloned by us [Wang et al., 1989]. FGF1 is expressed in ventral cochlear neurons, olfactory bulbs and hippocampal neurons but not in glial cells [Alam et al., 1996]. The brain-specific FGF-1B promoter is active only in the brain [Chiu et al., 2001; Chiu et al., 2000]. Interestingly, it has been shown that FGF-1B mRNA is upregulated for the maintenance of NSCs in hippocampus dentate gyrus in response to activity-induced neurogenesis [Ma et al., 2009]. Furthermore, FGF-1B promoter (-540 to +31)-driven GFP reporter (F1B-GFP) could be used to isolate NSCs with self-renewal and multipotent capacities from human glioblastoma tissues [Hsu et al., 2009b], and developing, neonatal or adult mouse brains [Hsu et al., 2009b; Lee et al., 2009]. We have developed a series of patented technology in which NSCs could

be isolated as GFP positive cells when adult mouse brain cells were transfected with F1B-GFP plasmid (USA patent No. 6,984,518; 7,045,678; and 7,745,214). This F1B-GFP plasmid comprises the GFP coding sequences driven by the human FGF1 promoter [Hsu et al., 2009b]. Notably, F1BGFP-selected NSPCs from mouse brains were able to repair the damaged sciatic nerve of paraplegic rats [Hsu et al., 2009a; Lin et al., 2008].

Personalized Regenerative Medicine: Patient-specific Neural Stem Cells and Functional NeuronsThe promising cell sources for personalized regenerative medicine is from patients’ skin fibroblasts through transcription factors-mediated reprogramming, such as induced pluripotent stem (iPS) cells [Takahashi et al., 2007]. Development of iPS cells from patients with amyotrophic lateral sclerosis further attests the possibility of future cell-based

therapy [Dimos et al., 2008]. However, recent studies using iPS cells have shown sizeable genetic and epigenetic abnormalities in iPS cells [Pera, 2011]. Recently, Vierbuchen et al. [Vierbuchen et al., 2010] found that a combination of just three factors (Ascl1, Brn2 and either Myt1l or Zic1) was sufficient to convert fibroblasts into neurons (iN). These iN cells expressed a variety of neuronal markers and were capable of firing action potentials. Furthermore, when cultured with mouse neural cells, the iN cells received both excitatory and inhibitory synaptic connections from the mouse neurons, and were able to form functional synapses with each other. Ascl1 alone was able to produce cells with immature neuronal features, but co-infection with Brn2 and Zic1 was required to produce cells with more mature neuronal features. In the past six months, six different laboratories independently demonstrated that adult human fibroblasts from a skin biopsy could be efficiently converted into functional neurons [Yang et al., 2011]. Although more accessible cell types such as peripheral blood could turn into iPS cells [Yamanaka, 2010], whether blood could be induced to neurons is yet unknown. In addition, transplantation

Table 1. Current Clinical Trials of Human Neural Stem Cells and Embryonic Stem Cells

COMPANY/ORGANIZATION

CELL PRODUCT DISEASECLINICAL TRIAL

STATUS

StemCells Inc., CA HuCNS-SC® (fetal derived human NSCs) Spinal cord injury Phase I/II

NeuroGeneration, CA Autologous NSC-derived Neurons Advanced PD Phase I – completed

Phase II – clinical hold

Neuralstem Inc., MD Fetal derived spinal cord SCs ALS Phase I

ReNeuron, UK ReN001 Immortalized huNSCs Stroke Phase I

City of Hope, CA HB1.F3.CD Immortalized hu NSCs Glioma Phase I

Geron Inc. Human ESC-derived oligodendrocyte progenitor cells (GRNOPC1)

Complete subacute thoracic spinal cord injuries. T3 to T10 segments between seven and 14 days after injury

Phase I

Advanced Cell Technologies Retinal Pigment Epithelium derived from human ESCs.

Stargardt’s Macular Dystrophy (juvenile macular degeneration)

Phase I/II

Age-related Macular Degeneration Phase I/II

California Stem CellHuman motor neuron progenitor cells derived from human ESCs

Spinal muscular atrophyType 1 Phase I: Currently on hold 2011

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Anderson L, Burnstein RM, He X, Luce R, Furlong R, Foltynie T, Sykacek P, Menon DK, Caldwell MA. 2007. Gene expression changes in long term expanded human neural progenitor cells passaged by chopping lead to loss of neurogenic potential in vivo. Experimental neurology 204:512-524.

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STUDY INJURY TYPE/ANIMAL MODEL CELLULAR TYPE/FACTOR MAJOR FINDINGS

Dezawa et al. (2001)Sciatic nerve injury in rats(1.5 cm gap)

Bone marrow MSCs differentiated into Schwann-like cells suspended in Matrigel

Successful nerve regeneration and myelination

injected into hollow fibers

McKenzie et al. (2006)Sciatic nerve crush injury inmyelin-deficient mice

Skin-derived precursors differentiated into Schwann cells

Remyelination and functional recovery

Marchesi et al. (2007)Sciatic nerve injury in rats(1.6 cm gap)

Guides filled with skin-derived stem cells

Functional recovery and myelination

Cui et al (2008)Sciatic nerve injury in rats(1.0 cm gap)

Mouse ES cell-derived neural progenitor cells

Remyelination and functional recovery

Lin et al., 2008, Hsu et al. (2009a),

Sciatic nerve injury in rats(1.0 cm gap)

Nerve conduit seeded with F1BGFP(+) mouse neural stem cells Axon regeneration

Chen et al. (2010)Acutely distracted sciatic nerves in rabbits

Neural stem cellsFunctional recovery and nerve regeneration

Gu et al. (2010)Sciatic nerve injury in rats(1.0 cm gap)

Rat fetal neural stem cells

Transplanted neural stem cells differentiated into neurons in peripheral nerves that synthesize and secreted synaptophysin

Cheng et al. (2010) Sciatic nerve crush injury in ratshuman amniotic fluid-derived mesenchymal stem cells and

Functional recovery and nerve regeneration

Di Summa et al. (2010)Sciatic nerve injury in rats(1.0 cm gap)

Nerve fibrin conduits seeded with adipose derived stem cells

Enhanced peripheral nerve repair

Reid et al. (2011)Sciatic nerve injury in rats(1.0 cm gap)

Adipose derived stem cellsDorsal root ganglia protection from apoptosis

Table 2. Preclinical Studies Evaluating Stem Cell Therapy for Peripheral Nerve Repair

experiments will be necessary to see whether iN cells can integrate into the damaged tissues and ameliorate disease in animal models [Nicholas and Kriegstein, 2010]. A way of expanding cell numbers is also required. Thus, it is expected to know that reprogramming allows the conversion of mouse fibroblast to induced NSCs. It is also

noted that FGFs have been shown to be

essential in culturing ESCs and NSCs. FGF1 has been shown to regulate cell proliferation, cell division and neurogenesis. Cheng et al. [Cheng et al., 1996] and Lee et al. [Lee et al., 2002] showed that the combination of FGF1 treatment and peripheral nerve grafts could restore hind limb function in adult paraplegic rats. More recently, it was shown that similar

treatments benefited the patients with

common peroneal nerve lesions as well [Tsai et al., 2009]. Thus, in addition to the stem cell-based therapy, direct clinical application of FGF1 will likely benefit the patients. Given the significance of FGF1 in the treatment of spinal cord injury and PNI, future efforts generating FGF1-expressing NSCs or iN will have promising potential in regenerative medicine and in the treatments of central and peripheral nerve system diseases.

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Uchida N, Buck DW, He D, Reitsma MJ, Masek M, Phan TV, Tsukamoto AS, Gage FH, Weissman IL. 2000. Direct isolation of human central nervous system stem cells. Proceedings of the National Academy of Sciences of the United States of America 97:14720-14725.

Vierbuchen T, Ostermeier A, Pang ZP, Kokubu Y, Sudhof TC, Wernig M. 2010. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463:1035-1041.

Wang WP, Lehtoma K, Varban ML, Krishnan I and Chiu IM. 1989. Cloning of the gene coding for human class 1 heparin-binding growth factor and its expression in fetal tissues. Molecular and cellular biology 9:2387-2395.

Yamanaka S. 2010. Patient-specific pluripotent stem cells become even more accessible. Cell stem cell 7:1-2.

Yang N, Ng YH, Pang ZP, Sudhof TC, Wernig M. 2011. Induced neuronal cells: how to make and define a neuron. Cell stem cell 9:517-525.

Zlokovic BV. 2011. Neurovascular pathways to neurodegeneration in Alzheimer’s disease and other disorders. Nature reviews Neuroscience 12:723-738.

Dr. Ing-Ming Chiu is a Distinguished Investigator and Director of Division of Regenerative Medicine in Institute of Cellular and System Medicine, National Health Research Institute in Taiwan. Dr. Chiu has focused his interests in three areas: (1) Therapeutic application and basic biology of neural stem cells and embryonic stem cells in neural degenerative disorders and neural repair, (2) The role of FGF1 in development and human diseases, and (3) Transcriptional regulation of FGF1. He has more than 100 publications and was granted nine patents with four more pending approval. In 2010, he was elected a fellow of American Association for the Advancement of Science. He was also awarded “National Innovation Award” in the Academic Research Category in 2011.

Dr. Yi-Chao Hsu is a postdoctoral fellow in Dr. Ing-Ming Chiu’s laboratory at the National Health Research Institutes, Taiwan. He received his Ph.D. in Pharmacology from National Yang-Ming University, Taiwan.

About the Authors

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S tem ce l l s a r e an eme rgen t biotechnology with the potential to generate enormous benefits in the

treatment of significant human diseases and conditions. They are currently used routinely in the treatment of certain types of blood and immune disorders and new approaches are currently being investigated in Phase I/II clinical trials. These approaches include the use of bone marrow-derived stem cells for myocardial infarcts,1 spinal cord injury,2 multiple sclerosis,3 and Parkinson’s disease.4 In addition, four separate trials using products derived from human embryonic stem cells (ESC),5,6 somatic ‘adult’ stem cells (ASC),7 and induced-pluripotent stem cells8 for the treatment of dry age-related macular degeneration are either underway or are expected to commence in near future.

However, these exciting developments are being overshadowed by an alarming number of clinics claiming to offer treatments with stem cells without having either been approved for use by regulatory authorities or demonstrated to be safe or effective.9 These clinics typically market stem cells directly to the consumer via the Internet for an unlikely range of conditions and illnesses.10,11 The interventions often come with hefty price tags and can be particularly expensive where overseas travel is required. Many are provided in countries with weak regulatory infrastructures, such as Mexico, China and Russia, although reports have also emerged of physicians offering them in the United States, the European Union and parts of South East Asia.

In this article, I report on some of the regulatory challenges being raised in an ongoing law suit between the Food and Drug Administration (FDA) and a Colorado-based company that is marketing an unapproved autologous ASC product in the US. I then show what the Governor of Texas and the Texas Medical Board have in common with a Korean stem cell company implicated in the deaths of two patients. I will then comment on why these events should be considered a major concern for the biotech industry as whole and what should be done about it. First, however, I will briefly describe the regulatory context of stem cell-based products.


Tamra LysaghtNational University of Singapore, Singapore

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The Regulatory Context From a regulatory perspective, stem cells are human cells and tissue-based products (HCT/Ps). In most developed countries, HCT/Ps are regulated by authorities that oversee interstate commerce in drugs, biologics and devices. In the US, this task is delegated to the FDA. As elsewhere, unless an HCT/P qualifies for an exemption, FDA approval is required before it can be sold commercially across interstate borders. Exemptions are generally permitted only if the HCT/P is minimally manipulated, intended for homologous use, is not combined with another article (except for water, sterilizing, preservation or storage agents), and either has no systemic or metabolic effect, or is for autologous use, allogeneic use in first-second degree blood relative or reproductive use.

HCT/Ps that do not meet all of these criteria, and have not yet been approved for marketing, may be only administered under an Investigational New Drug (IND) application. INDs must be accompanied with preclinical data, including animal pharmacology and toxicology studies, and detailed protocols for proposed clinical studies that have been approved by an Institutional Review Board (IRB). Only after the safety and efficacy of a product is demonstrated, which is typically done through formal clinical trials, will a manufacturer of an IND be issued with a valid biologics license to market the product. Manufacturers must then register with the regulator and conform to current Good Manufacturing Practices (cGMP).

The Legal DisputeThe legal dispute between the FDA and the Colorado-based company named Regenerative Sciences, Inc. began in 2008, when the regulator issued an ‘untitled letter’ to its medical director, Christopher Centino. In this letter, the FDA claimed that the autologous mesenchymal stem cells (MSCs) being marketed on the company’s website for musculoskeletal and spinal injuries were HCT/Ps that required a valid biologics licence.12 As neither a valid licence

nor IND was in effect, the company was asked to cease being in violation of the federal regulations. Regenerative Sciences responded by denying that their MSCs are drugs or biological products and, therefore, should not fall under the regulatory authority of the FDA.

In the legal proceedings that have followed, which are described in more detail elsewhere,13 the FDA has claimed that the MSCs used in Regenerative Science’s procedure are an adulterated biological drug product that should be licenced, branded and manufactured according to cGMP. The company has denied this claim and instead argues that their procedure constitutes the practice of medicine, over which the FDA has no authority. Medical procedures are instead regulated by professional practice standards, state licensing boards, professional accreditation bodies, third party payers, and medical malpractice laws.14 Thus, if the FDA injunction does not hold, it could potentially pave the way for virtually anyone with a medical licence to culture autologous ASCs and inject them back into patients, with little to no regulatory oversight.

From Texas to South Korea (and back)This outcome, it appears, would be welcomed by some. In 2011, the Governor of Texas and then-presidential candidate for the Republican Party, Rick Perry, was reported to have been injected with his own cultivated stem cells for a back complaint.15 The procedure was reportedly carried out at an undisclosed location in Texas by Houston orthopedic surgeon, and Perry’s personal friend, Dr Stanley Jones.16 Jones had apparently never performed the procedure but had received a similar treatment in Japan for his arthritis. The autologous MSCs used in both procedures were isolated and grown by a South Korean company called RNL Bio.

A month prior to Perry’s procedure, it was reported that Texas lawmakers had passed a health care bill that authorized the creation of a state ASC bank and that the Governor’s office had been in discussions with the Texas Medical Board (TMB) about regulating the procedure.17 The TMB responded by

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announcing that it would look to provide oversight of stem cell transplants as a medical procedure with new rules to guide physicians in their use of investigative agents such as stem cells.18 According to the draft guidelines, physicians may use unapproved stem cell products providing they first gain approval from an accredited IRB and follow state and federal laws concerning the use of therapeutic agents, which include the FDA regulations on HTC/Ps.

Key industry players have been accused of lobbying the TMB heavily not to introduce onerous regulations on physicians. One of those is a company called Celltex Therapeutics Corp, which was co-founded by Stanley Jones and a financial supporter of Rick Perry. Last year, prior to the enactment of the Texas legislation, Celltex signed a reported $30 million licensing agreement with RNL Bio for the exclusive rights to market their stem cell technology in North America.19 RNL Bio, which is based in Seoul, and is better known for its commercial dog cloning services, came under investigation in 2010 following the deaths of two Korean patients who had received injections of the company’s stem cells in Thailand and Japan.20

While the formal investigation of RNL Bio continues, the International Cellular Medicine Society (ICMS) conducted one of its own in 2010. The ICMS is a US-based

organisation that was co-founded by the medical director of Regenerative Sciences, the company currently in court with the FDA. Assisting with their evaluation of RNL Bio’s ethical and clinical practices was Glenn McGee, Editor-in-Chief of the American Journal of Bioethics (AJOB) at the time. The ICMS investigation found that one of the deaths were likely caused by the stem cell procedure.21 RNL Bio has since been expelled from the ICMS membership for failing to comply with the recommendations made in its investigation.22 In a final twist, McGee resigned from his post at AJOB in late 2011 to take up a senior position at Celltex, but reportedly quit the job in February 2012 amidst much controversy over the potential for conflicts of interest at the journal.23

ImplicationsAs these events continue to unfold, there are some serious concerns that should alarm the biotech industry more widely. The first relates to the lack of published data demonstrating that stem cells are safe or efficacious to use outside of their existing applications in certain blood and immune disorders. Enthusiasm to be first to market can sometimes override good judgment when novel treatments are pushed into the clinic

prematurely and before for their safety and effectiveness can be properly assessed. Few examples remind us of this more starkly than when bone marrow transplant was used with high dose chemotherapy for the treatment of breast cancer.24 This so-called therapy was eventually shown to be no more effective than the standard treatment, but not before substantial amounts of public resources and private investment were wasted commercializing an intervention that simply did not work.

A second concern is the potential for the loss of credibility and loss of public trust that the industry may face if, in the near future, these unapproved interventions fail to live up to the hype and expectations being encouraged by their proponents. For an industry that struggles against authorities in some jurisdictions, such as the EU which has previously imposed trade restrictions on genetically engineered products25 and banned patents on ESCs,26 it may be timely to discuss the options for self-regulation. For, it is conceivable that if the industry does not implement guidelines and sanctions of its own, then the regulatory authorities, especially those in the major markets of the US and EU, most certainly will and it is unlikely that they will be aimed at protecting commercial interests as a first priority.

References1. Bartunek J, Vanderheyden M, Hill J, Terzic A. Cells as biologics for cardiac repair in ischaemic heart failure. Heart. May 1, 2010 2010;96(10):792-800.

2. Rossi SL, Keirstead HS. Stem cells and spinal cord regeneration. Current Opinion in Biotechnology. 2009;20(5):552-562.

3. Pasquini MC, Griffith LM, Arnold DL, et al. Hematopoietic Stem Cell Transplantation for Multiple Sclerosis: Collaboration of the CIBMTR and EBMT to Facilitate International Clinical Studies. Biology of Blood and Marrow Transplantation. 2010;16(8):1076-1083.

4. Wijeyekoon R, Barker RA. Cell replacement therapy for Parkinson’s disease. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2009;1792(7):688-702.

5. Advanced Cell Technology. Advanced Cell Technology Receives FDA Clearance For Clinical Trials Using Embryonic Stem Cells to Treat Age-Related Macular Degeneration. 2011; http://www.advancedcell.com/news-and-media/press-releases/advanced-cell-technology- receives-fda-clearance-for-clinical-trials-using-embryonic-stem-cells-to-tre/. Accessed 7 January 2011.

6. London Project. Taking the Therapy to Clinical Trials. 2011; http://www.thelondonproject.org/OurVision/TheProject/?id=1432. Accessed 21 Feb 2012.

7. StemCells Inc. News Release: StemCells, Inc. Receives FDA Authorization for Age-Related Macular Degeneration Clinical Trial. 2012; http://investor.stemcellsinc.com/phoenix.zhtml?c=86230&p=irol-newsArticle_print&ID=1655922&highlight=. Accessed 21 Feb 2012.

8. Waters H. An eye for stem cells. The Scientist 2011; http://the-scientist.com/2011/12/01/an-eye-for-stem-cells/. Accessed 13 Dec 2011.

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Dr Tamra Lysaght's research interests lie broadly around the ethical, sociopolitical and regulatory issues surrounding stem cell science and the clinical translation of regenerative medicines. She is currently a Research Fellow at the Centre for Biomedical Ethics at the National University of Singapore where she is developing a research program into the ethics of innovative stem cell technologies and regulatory policies for stem cell science in the Asia Pacific. In the last two years, she has worked with the Ethics Committee of the Human Genome Organisation and the WHO Technical Working Group on Ethics, and also forms part of the Ethics Core of the Translational Clinical Research Programme at the Institute of Mental Health in Singapore.

About the Author

9. Lau D, Ogbogu U, Taylor B, Stafinski T, Menon D, Caulfield T. Stem Cell Clinics Online: The Direct-to-Consumer Portrayal of Stem Cell Medicine. Cell Stem Cell. 2008;3(6):591-594.

10. Regenberg AC, Hutchinson LA, Schanker B, Mathews DJH. Medicine on the Fringe: Stem Cell-Based Interventions in Advance of Evidence. Stem Cells. 2009;27(9):2312-2319.

11. Kiatpongsan S, Sipp D. Monitoring and Regulating Offshore Stem Cell Clinics. Science. March 20, 2009 2009;323(5921):1564-1565.

12. Food and Drug Administration. Vaccines, blood and biologics: Regenerative Sciences Inc. 2008; http://www.fda.gov/ BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/ComplianceActivities/Enforcement/UntitledLetters/ucm091991. htm. Accessed 18 November 2010.

13. Lysaght T, Campbell AV. Regulating Autologous Adult Stem Cells: The FDA Steps Up. Cell Stem Cell. 2011;9(5):393-396.

14. Taylor PL. Overseeing innovative therapy without mistaking it for research: a function-based model based on old truths, new capacities, and lessons from stem cells. J. Law Med. Ethics. Summer 2010;38(2):286-302.

15. Cyranoski D. Texas prepares to fight for stem cells. Nature. 2011;477:377-378.

16. Ramshaw E. Perry’s adult stem cell treatment was doctor’s first attempt. 2011; http://www.texastribune.org/texas-people/rick-perry/ perrys-stem-cell-treatment-was-doctors-first-attem/. Accessed 22 Feb 2012.

17. Ramshaw E. Perry, allies lay groundwork for TX stem cell industry. 2011; http://www.texastribune.org/texas-people/rick-perry/perry- allies-lay-groundwork-tx-stem-cell-industry/. Accessed 24 August 2011.

18. Roser MA. Texas Medical Board seeks to regulate stem cell transplants, like Perry’s back treatment. 2011; 26 August 2011:http://www. statesman.com/news/texas-politics/texas-medical-board-seeks-to-regulate-stem-cell-1796362.html. Accessed 1 September 2011.

19. Isikoff M. Rick Perry pushed bill that could benefit stem cell doctor’s firm 2011; http://www.msnbc.msn.com/id/44291973/ns/politics- decision_2012/t/rick-perry-pushed-bill-could-benefit-stem-cell-doctors-firm/#.T0R8w1Euh8E. Accessed 22 Feb 2012.

20. Cyranoski D. Korean deaths spark inquiry. Nature. 2010;468:485.

21. International Cellular Medicine Society. ICMS announces investigation findings. 2010; http://www.cellmedicinesociety.org/home/ news/latest/317-icms-announces-investigation-findings. Accessed 28 February 2012.

22. Freeman M, Audley D. International Cell Medicine Society Releases Statement on RNL. 2011; http://msnbcmedia.msn.com/i/MSNBC/ Sections/TVNews/Nightly%20News/2011%20Stories/Perry_Celltex_PDFs/4_Stem%20cell%20society%20statement.pdf. Accessed 24 Feb 2012.

23. Cyranoski D. Editor’s move sparks backlash. Nature. 2012;482(7386):449–450.

24. Rettig RA, Jacobson PD, Farquhar CM, Aubry WM. False Hope: Bone Marrow Transplantation for Breast Cancer New York: Oxford University Press; 2007.

25. McCabe H, Butler D. European Union tightens GMO regulations. Nature. 1999;400(6739):7.

26. Callaway E. European court bans patents based on embryonic stem cells. Nature. 2011;Published online 18 October 2011:doi:10.1038/ news.2011.1597

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C o-organised by the Hong Kong Trade Development Council (HKTDC) and The Modernized Chinese Medicine

International Association Limited (MCMIA), the International Conference & Exhibition of the Modernization of Chinese Medicine & Health Products (ICMCM) was held at the Hong Kong Convention and Exhibition Centre (HKCEC) from 11 to 15 August 2011. The event served as a perfect platform for practitioners and industry players to meet and engage in business, and allowed visitors from around the world to have a better understanding of Chinese medicine.

Entering into its tenth year, the exhibition showcased a wide range of products and services, ranging from Chinese medicine, functional food and products, health supplement, health care and therapy, to raw materials, equipment and research & development. The fair welcomed 138 exhibitors not only from traditional markets of Chinese medicine such as Hong Kong, Macau, Chinese mainland and Taiwan, but also from Japan, Malaysia, Singapore and Canada. Group pavilions were set up to represent Guizhou, Heilongjiang, Jiangsu, Jilin, Qinghai and Sichuan to showcase the

Annual Pivotal Event for Global Chinese Medicine Industry

top-quality products from these provinces of the Chinese mainland. The Chinese mainland was once again the major origin of exhibitors, indicating its leading position in the market.

The five-day event attracted a total of 9,365 trade buyers, with 3,404 of them coming from overseas. The top ten countries and regions visiting were the Chinese mainland, Japan, the Philippines, Korea, Taiwan, Malaysia, Singapore, Canada, Macau and Thailand.

Furthermore, 50 buying missions representing over 1,200 companies were organised from 20 countries & regions, including top importers l ike Apollo Enterprises Ltd from India and Sheng Chang Pharmaceutical Co. Ltd. from Taiwan.

The first two days of the exhibition were dedicated to trade, allowing manufacturers, dealers, trade buyers, researchers and industry professionals to exchange ideas and establish business contacts. The exhibition was open to the public on the remaining three days (August 13 to 15). Exhibitors were delighted to present their latest inventions, knowledge and products to end users and collect comments from them. Exhibitors’

Forum and Public Forum of Chinese Medicine Health - “Healthy Life in the Wisdom of Chinese Medicine” were organized to provide the general public with useful information on health related issues.

Professional ConferenceAt the heart of the event was the 10th ICMCM Conference - “Milestones in Chinese Medicine Development” held on August 11 and 12. Top-notch speakers from Canada, the Chinese mainland, Israel, Japan, Taiwan, Thailand and Hong Kong were invited to share their valuable insights on topics such as “Strategies for International Business Development” and “Regulatory Harmonization and Anti-viral & Neuro Treatments”.

Unparallel SynergyThe ICMCM was complemented by the concurrent Food Expo and Hong Kong International Tea Fair, which provided buyers with additional sourcing synergy to seek exciting new business ventures and partners, and gave general public the chance to visit three top-quality exhibitions in one go.

A Heads-up EventThe ICMCM is an annual pivotal event for the global Chinese medicine industry that cannot be missed. The next edition will be held from August 16 to 18, 2012, together with the Food Expo and Hong Kong International Tea Fair.

For further information, please visit the official website of ICMCM at http://www.hktdc.com/icmcm/

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India Unveils Roadmap for Cooperation with Africa

T he 1st India-Africa S&T Ministers’ Conference and Tech Expo got off to a business-like start recently with Shri

Vilasrao Deshmukh, Minister for Science & Technology and Earth Sciences, reiterating the commitments of Government of India to the declarations of India-Africa Summits and outlined a five-fold agenda to pave the way for a concrete roadmap of cooperation in science, technology and innovation with African countries.

The two-day Conference and Tech Expo was organized jointly by the Ministry of Science and Technology (Department of Science & Technology) and Ministry of External Affairs (Government of India) and FICCI.

The Minister said that facilitation of India-Africa interaction at a higher level could built around the following five areas:

• Visits of expert delegations from India to African countries for identification of an initial set of potential sectors for collaboration and technology transfer;

• Organization of joint workshops, technology expos and road shows for requirement assessment and feasibility studies;

• Identification of agencies that can provide technologies for transfer to African countries;

• Development of appropriate linkages between industry associations and SMEs; and

• Facilitating technology transfer and establishment of pilot scale demonstration plants in Africa

The Minister observed that the Indian IT sector is in a growth phase. The infrastructure of our laboratories is being strengthened. Most of India’s R&D and academic institutions are now actively engaged with international collaborations which allow mobility of students and researchers.

“I see here an opportunity for science and engineering students at the Masters and PhD levels from Africa to undertake student internships in Indian research

institutions. Through this exposure we can connect young scientists and build bridges between next generations. While brain drain to other developed countries may be an issue for developing economies, I can assure you that while sending your students to India such concerns may not be relevant,” Mr. Deshmukh said.

He said technology transfer and adoption is the central theme of the India-Africa S&T partnership . “Some for the Indian home grown technologies and innovations which are frugal and affordable could easily touch the lives of the untouched people in both Indian and Africa. Affordable innovations and traditional knowledge sectors could form the wings of Indo-Africa partnership,” he said.

Speaking on the occasion, Ms. Preneet Kaur, Minister of State for External Affairs, stated, “In a demonstration of India’s continued commitment in Africa, India has successfully implemented the pan-African e-Network Project, including tele-education, tele-medicine and connectivity between leaders in 47 African countries. An agreement has also been signed for its implementation in the 48th country, South Sudan, recently.

Dr. (Ms.) Nadia Eskander Zakhary, Minister of Scientific Research of the Arab Republic of Egypt and Chair of the African Ministerial Conference on Science and Technology, informed the delegates that 2010-20 has been declared as the “Decade for Africa’. “The challenge for the African nations was to plan, unite and engage in strategic efforts for the betterment of the people,” she said and urged her Indian colleagues to support Africa’s S&T initiatives for the sake of future generations.

Dr. Ashwani Kumar, Minister of State for Science & Technology and Planning, Government of India, pointed out that India remains committed capacity building in the HRD sector in Africa. “India”, he said, “proposes to establish several new institutions at the pan-African level, including an India-Africa Food Processing Cluster, India-Africa Integrated Textiles Cluster, India-Africa Centre for Medium

Range Weather Forecasting.” The latter, she said, would harness satellite technology for the agriculture and fisheries sectors as well as contribute towards disaster preparedness and management of natural resources. “We have received a request to support the establishment of an India-pan-Africa University for Life and Earth Sciences in Nigeria and would be happy to support this important venture,” he added.

Prof.JeanPierreOEzin,Commissioner,Human Resources, Science & Technology, African Union Commission, in his address, said that science and technology was today an important component of the political agenda in Africa. He expressed confidence that the conference would help create a long term strategic partnership in ways that would touch the lives of the people in the two regions.

Shri R. V. Kanoria, President, FICCI, informed the delegates that discussions were at an advanced stage with some African nations to launch the DRD-FICCI technology commercialization programmes in Africa. The proposed programmes, notably in Senegal and Rwanda, aim at taking relevant Indian technologies to Africa with the mutually beneficial goal of strengthening industry while providing new international markets for Indian innovations.

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S hri Sharad Pawar, Minister of Agriculture and Food Processing Industries, India, has stressed the need

for according high priority to soil health and nutrient management for sustainable farm growth. Addressing the National Kharif Conference 2012, Shri Pawar said that soil health, which is crucial for ensuring farm productivity, has been witnessing a declining trend over the years.

The Minister said, “Adoption of multi-nutrient carriers that are soil and crop specific and customized on the basis of soil testing is emerging as a viable alternative to conventional approach. Besides, organic nutrient sources like farm yard manure, crop residues, vermi compost, bio fertilizers, green manure etc. can also play a key role in adopting eco-friendly agriculture.”

Indian Agriculture Minister Calls For Improved Soil HealthOnthenewinitiativeslaunchedbythe

Government to enhance farm productivity in Eastern India and rainfed agriculture, the Minister said, “while Bringing Green Revolution to the Eastern India (BGREI) scheme aims at improving productivity of rice based cropping system, Rainfed Area Development Programme (RADP) promotes customized and focused interventions to rainfed areas for enhancing food and livelihood security through integrated farming. We will continue with our endeavour this year too.”

Referring to shortage of farm labour, Shri Pawar said that he is laying enhanced emphasis on farm mechanization with focus on small and marginal farmers. He said that his Ministry is proposing a large programme for agricultural mechanization during the

12th Plan along with an attempt to utilize Mahatma Gandhi National Rural Employment Guarantee Act for augmenting activities that directly add to farm productivity.

The Minister informed the Conference that the Rastriya Krishi Vikas Yojana (RKVY) which has emerged as the principal vehicle of financing the development programmes and infrastructure investment in States in agriculture and allied sectors will be revisited in 12th Plan to give more focus on development of agri-infrastructure.

Sh r i Pawar u rged the S ta tes to formulate State specific Agriculture Infrastructure Development Plan and work on Agriculture Reforms Policy for boosting development of agriculture and allied sector taking into account their respective agro-climatic conditions.

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G E Global Research, the central technology development arm for GE Healthcare and all of GE’s

businesses, has signed a Memorandum of Understanding (MOU) with Singapore’sAgency of Science, Technology and Research (A*STAR). This agreement will focus on advancing current medical imaging technologies and diagnostics to enable more accurate, earlier and faster clinical diagnoses of cancer and other diseases. The partnership between A*STAR and GE Global Research brings together two world-class research institutions, integrating their deep domain expertise in biomedical, science, and engineering capabilities to support this effort. ThisMOUexpandsuponaproductivecollaboration between GE and A*STAR’s S ingapore B io imaging Consort ium (SBIC) using Hyperpolarized Carbon-13 technology. Early results exploring sub second bio-chemical imaging in Oncologyapplications helped pave the way for a broader scientific collaboration on projects in medical diagnostics and medical imaging. The goal is to improve diagnosis and tissue characterization in diseases that

A*STAR, GE Global Research To Develop Integrated Advanced Medical Imaging

are prevalent in the Asian population, such as liver, lung, and gastric cancers. Michael Idelchik, Vice President of Advanced Technology Programs at GE Global Research, said, “To more effectively combat cancer and other deadly diseases, more advanced diagnostic tools will be needed to help doctors become more prescriptive in their diagnoses and treatment regimens. Combining A*STAR’s world-class biomedical and clinical expertise with GE’s strengths in diagnostic and molecular imaging, we have an exciting opportunity to take medical diagnosis to this next level. Specifically, A*STAR will help us address cancers and other diseases more common in Asia and where pathology and outcomes are different as compared to the rest of the world.” Professor Low Teck Seng, Managing Director of A*STAR, said, “This win-win public-private partnership between A*STAR and GE comes at an opportune time with the increasing research interest in diseases affecting the Asian population. I am confident that A*STAR’s cross-disciplinary capabilities in both the biomedical, and physical sciences & engineering research will

complement GE’s expertise in diagnostic and molecular imaging to meet today’s complex healthcare challenges and enhance lives.” As part of the MOU, A*STAR andGE Global Research will collaborate to enhance medical imaging technologies in imaging modalities, ranging from magnetic resonance imaging (MRI) and positron emission tomography (PET) to computed tomography (CT). In a Frost & Sullivan global market analysis report, the medical imaging sector was valued at about US$25 billion as of 2008, with MRI and CT scanners accounting for a combined 40% of the total global device medical imaging market. In one project, scientists from A*STAR’s Institute of Microelectronics (IME) and GE scientists will explore the development of new imaging technologies to improve the speed and accuracy of clinical cancer diagnosis. Leveraging IME’s network and partnerships with the microelectronics industry, this project could result in the development of a new local industry for Singapore in the healthcare technologies area. In another project, A*STAR’s Singapore Bioimaging Consortium (SBIC) and GE plan to develop novel imaging markers for hepatic cellular carcinoma (HCC), the most common type of liver cancer in Asia. This project will integrate biomedical imaging and pre-clinical model development expertise from SBIC with GE’s molecular diagnostics technology to develop innovative, proprietary platforms to help advance the unique characterization of HCC in each patient. In this manner, the goal is that a specific type of cancer would be identified and the therapy tailored to each patient. This project encompasses a range of medical diagnostic technologies from imaging to molecular pathology biomarkers appropriate to HCC, relevant to the Asian population. Building on a close partnership with local hospitals, success in this project may lead to accelerated and accurate cancer diagnosis that enables more prescriptive and effective cancer treatments for patients. This will support A*STAR’s efforts to develop Singapore as a Center for Oncology andMolecular Pathology.

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P RA, a leading Clinical Research Organization,continuesitsexpansion across the burgeoning Asia-Pacific

region with the recent opening of an office in Singapore. The office will serve as PRA’s central clinical operations location for the region, strengthening the quality and timeliness of its deliverables to sponsors.

The Singapore facility, conveniently located in the heart of the city-state, will provide monitoring, project management, safety reporting, feasibility and other services.

“Building our capability across the region with the center in Singapore permits

PRA Opens Office in Singaporeour sponsors to experience a wider range of services and expertise,” said Helen Neal, PRA’s Director of Clinical Operations forAsia-Pacific. “This location provides our teams with access to experts and resources within their time zone, positively impacting our speed and quality of delivery on projects.”

Geographically renowned as the region’s pharmaceutical and biomedical center, Singapore is the ideal location from which to deliver centralized study support to the region. This city-state serves as an excellent logistics hub with high fluency rates in English, and its government strongly

promotesandencouragesCROstoconductcutting-edge clinical research.

“OpeningaSingaporeofficeforPRAisan exciting and significant milestone in the company's evolution and is evidence that PRA acknowledges and embraces the full potential that the Asia-Pacific region can bring to the global business going forward,” said Ms. Neal.

PRA has been strategically expanding our Asia-Pacific operations since 2000. In addition to Singapore, PRA operates in eight other countries within the region: Korea, Taiwan, China, Hong Kong, Thailand, Australia, New Zealand, and India.

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T he Bristol-Myers Squibb Foundation has awarded three new grants to improve prevention, diagnosis and care

of hepatitis B (HBV) and hepatitis C (HCV) in China and India as part of its Delivering Hope™: Awareness, Prevention and Care umbrella program which is committed to reducing hepatitis-related health disparities in Asia. China and India together have an estimated 123 million people chronically infected with HBV and 59 million people chronically infected with HCV, accounting for almost 50 percent of all HBV and HCV infections worldwide.

The grant recipients, which range in scope from national and regional government health, charitable non-profit and advocacy organizations, were announced at the Asian Pacific Association for the Study of the Liver (APASL) 2012 Conference in Taipei, Taiwan, where leaders in the hepatology field gathered to promote scientific advancement and education in the Asia Pacific region. Organizationsandprojectsreceivingsupportinclude:

Bristol-Myers Squibb Announces Grants to Tackle Hepatitis B and Hepatitis C in Asia

• The Chinese Foundation for Hepatitis Prevention and Control (CFHPC), working in partnership with the Chinese Center for Disease Control and Prevention and Shanxi Center for Disease Control, will enhance HCV prevention, diagnosis, support and care through training of physicians and health providers at various levels, as well as patient and community outreach in the Shanxi Province.

• TheShanghaiCharityFoundation(China) will create a first of its kind program targeting high risk groups with disease management initiatives for HCV and HBV in Shanghai. This will include vaccinations, screenings and behavior change programs to strengthen prevention efforts.

• TheLiverFoundation,WestBengal(India) will establish and maintain an advocacy platform focused on empowerment of hepatitis patients, ensuring knowledge and awareness of their disease, rights and privileges, as well as access to care.

“The benefit of these organizations and programs lies in their ability to empower

people in local communities with knowledge about prevention, diagnosis and care of hepatitis C and hepatitis B,” said John Damonti, president, Bristol-Myers Squibb Foundation. “Through Delivering Hope, The Bristol-Myers Squibb Foundation continues to harness expertise and resources in community-based programs, and leverage those best practices to help others.”

The mission of the Bristol-Myers Squibb Foundation is to help reduce health disparities in communities where the need is greatest. Marking a decade of support, Delivering Hope has invested and initiated 38 program grants across Asia totaling more than $9.7 million USD, specifically 16 grants in mainland China, three in Taiwan, 15 in India and four in Japan. In keeping with the Foundation’s commitment to sharing lessons learned, funding recipients participated in a two-day conference to discuss tracking and reporting outcomes, impact and best practices. These reports will be shared with the HBV and HCV community to enhance the body of knowledge on hepatitis prevention, care and support.

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B urrill & Company has announced the opening of an office in Taiwan led by Marietta Wu, M.D., Ph.D., whom

the company has promoted to Managing Director, Taiwan.

“Taiwan is in a strategically important region offering tremendous opportunities. By investing there, we can broaden access to emerging markets in China and East Asia. At the same time, we offer companies in Taiwan a window on innovation in the West and can help them grow their companies into players on the global stage,” says G. Steven Burrill, CEOofBurrill&Company,adiversifiedglobalfinancial services firm focused on the life sciences industry. “We are pleased to have someone with Marietta’s depth of experience lead our effort there.”

Wu, who joined Burrill & Company’s venture group in 2006, will open and manage the office, serve as the principal point of

Burrill & Company Expands into Taiwan

contact for the venture group’s investors in Taiwan, and manage investments in Taiwan-based companies. At Burrill, Dr. Wu has led investments in companies and fundraising efforts in Taiwan. She also served as acting COO of Waterstone Pharmaceuticals, aBurrill portfolio company. Prior to joining Burrill, Dr. Wu held professional positions with Edwards Lifesciences and Eli Lilly. As Director of Strategy at Edwards, Dr. Wu was responsible for strategic planning in its

biologicsprogram.AtEliLillyOncologyandIntegrated Biology, she oversaw finance, business development, pipeline valuation, organizational restructuring, and outsourcing strategy. She also worked at Eli Lilly’s M&A Group and with Oncology DiscoveryResearch. Dr. Wu founded BioHorizon, a consultancy focused on value creation in the life sciences industry across the Pacific Rim. She advised a number of biotech start-ups and venture capital firms.

Dr. Wu’s scientific expertise centers on basic and clinical research in oncology and neuroscience. She was selected by Nobel Laureate Dr. Marshall Nirenberg as an IRTA Fellow at the National Institutes of Health. She received her M.D. from Shanghai Jiaotong University School of Medicine, a Ph.D. in Medical Sciences from MedicalCollegeofOhio,andanMBAfromthe University of Michigan Business School.

Medistem Inc. announced today initiation of joint efforts with the Chinese conglomerate, Shanghai

Jia Fu Medical Apparatus Inc, in developing the Endometrial Regenerative Cell (ERC) “universal donor” stem cell product for the Chinese market. The initial focus of the collaboration will be treatment of critical limb ischemia, an advanced form of peripheral artery disease. Medistem has previously received FDA clearance to begin a Phase I clinical trial using the ERC stem cells in this patient population. A scientific publication providing the rationale and supporting data for utilization of ERC in treatment of critical limb ischemia may be found at http://www.translational-medicine.com/content/pdf/1479-5876-6-45.pdf.

“Medistem has over 10 peer-reviewed scientific papers published with academic groups from China over the past 5 years. The Team is very eager to expand our collaborations with China not only in science, but now in product development,”

Medistem Initiates Collaboration With Shanghai Jia Fu Medical Apparatus

saidThomasIchim,CEOofMedistem.“Giventhat Shanghai Jia Fu Medical Apparatus Inc is actively working in the area of clinical translation of cellular therapy, we believe this group is an ideal partner for our entry into development of therapeutics for the Chinese population.

Shanghai Jia Fu Medical Apparatus Inc currently has commercialized autologous cytokine activated killer cell therapy for cancer, as well as autologous bone marrow for treatment of critical limb ischemia; the company is interested in exploring expansion into allogeneic “off the shelf” products. As part of the initial collaboration, Medistem will provide ERC stem cells for research use and Shanghai Jia Fu Medical Apparatus Inc will support ongoing experiments. All data generated will be provided to Medistem.

“This is an example of a 2 +2 = 8 synergy,” said Mr. Wei Zhang, CEO ofShanghai Jia Fu Medical Apparatus Inc. “By leveraging cutting edge technology developed by Medistem in the context

of our existing cell research and therapy infrastructure, we hope to accelerate Medistem’s development program, while at the same time bring new research to China.”

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A dvinus Therapeutics, a research- based pharmaceutical development company promoted by the TATA

Group, and SignalChem Pharmaceuticals, a British Columbia-based drug discovery company, have started working on 15 programs based on SignalChem’s proprietary Kinase platform for targeted therapies in oncology. Each program is being developed with corresponding diagnostics and tracers for patient selection and biomarkers for follow up offering the right treatment to the right patient at the right time. The companies have received a very high level of interest from pharma companies to license and collaborate on the programs and therefore see a rapid progression to the clinic.

Advinus, SignalChem Commence Collaboration On New Anti-Cancer Drugs

Dr. Rashmi Barbhaiya, CEO andMD of Advinus said– “This collaboration is an exemplary model of polycentric and globally networked innovation for enhancing probability of success, reducing timelines, and focusing on cost-effective partnerships to bring innovative medicines to the market. This collaboration would also leverage leading edge biomedical research of British Columbia research institutions and Advinus’s capabilities and infrastructure to translate biomedical research discoveries into potential life-saving drugs.”

Dr. Jasbinder Sanghera, CEO ofSignalChem said – “This is a path-breaking partnership that leverages SignalChem's extensive biology and kinase expertise

and Advinus's end-to-end capabilities of medicinal chemistry, lead optimization, preclinical development and early-clinical development capabilities to rapidly and efficiently advance new anti-cancer drug candidates into human clinical trials.”

The collaboration was announced by B.C. Premier Christy Clark during her visit to India who said – “This is what our Jobs Trade Mission is all about – helping local companies make connections that will allow them to do business in India. We have an innovative technology and life sciences sector in B.C., which has a lot to offer India to support its economic growth. The partnership between Advinus and SignalChem will benefit people around the world.”

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M icroTissues, Inc. announced today that its 3D Petri Dish™is targeted towards helping to reduce the

numbers of animals used in research. The 3D Petri Dish™ is a new tool for the worldwide industry of life sciences research and drug discovery that grows living cells in three dimensions (3D). These 3D microtissues replicate the function of natural tissues and organs better than conventional 2D methods and are increasingly being used in toxicity testing of new drugs and cosmetics.

“Worldwide, efforts are underway to reduce the use of animals in research and we’re excited to be offering a new technology for toxicity testing” said Brian Morgan, Marketing Manager of MicroTissues, Inc. “The 3D Petri Dish™ is a reliable 3D cell culture technology that accurately produces natural cell-to-cell interactions and is easy to use. And, it forms 3D microtissues from human cells, so toxicity testing data is more relevant.”

Effective 2009, the European Union banned the use of animal testing for

3D Petri Dish to Reduce Animal Use in Researchcosmetic products and many believe the trend to reduce the use of animals in research will continue worldwide. MicroTissues, Inc. is helping to address this issue by offering eight products that are precision micro-molds used to cast 3D Petri Dishes™ that fit in standard multi-well plates. The 3D Petri Dish™ forms hundreds of multi-cellular 3D spheroids from cells useful for toxicity testing including hepatospheres, cardiospheres, mammospheres, neurospheres, and embryoid bodies. The 3D Petri Dish™ technology also forms microtissues with complex shapes having geometries that mimic natural organs. Over thirty different cell types,including primary human cells, have been shown to form 3D microtissues in the 3D Petri Dish™.

MicroTissues, Inc. a privately held company with an exclusive worldwide license to US and international patent applications on the 3D Petri Dish™, is advancing technologies and applications of 3D cell culture. The company’s products stand above the rest because they are

designed to create more natural and more reliable 3D cell culture environments based on cell-to-cell interactions in convenient and consistent formats that generate high content information. The company’s lead line of products, the 3D Petri Dish™, is serving the needs of researchers in a wide range of areas including cancer research, stem cell biology, toxicity testing, developmental biology, drug discovery, regenerative medicine and tissue engineering. In addition to products for basic research, MicroTissues, Inc. is using its platform technology to pursue applications in drug discovery and cell therapy.

V ermillion, Inc. (NASDAQ: VRML), a leading molecular diagnostics company, has received a notice of

allowance from the United States Patent andTrademarkOfficeforapatent,"Plateletbiomarkersforcancer."

The patent resulted from a collaboration with the late Dr. Judah Folkman, a renowned cancer expert, and identifies three biomarkers that can be used to assess changes in endogenous angiogenesis in a subject. Angiogenesis is commonly associated with cancer, and novel therapeutics such as bevacizumab (Avastin®) target angiogenesis to limit tumor recruitment of blood vessels.

The patented biomarkers, which are associated with platelets, can be used to

Vermillion Receives Patent on Platelet Biomarkers of Angiogenesis

measure ongoing angiogenic activity. The patent covers the measurement of these biomarkers over time and correlating changes in expression with the changing level of endogenous angiogenic activity. Consequently, this patent also enables the use of these biomarkers to monitor efficacy of therapy directed at angiogenic pathways.

"Vermillion is committed to thecreation of innovative, effective tests for cancerdiagnosisandpatientmanagement,"said Dr. Donald Munroe, the company's chief scientific officer. "Webelieve the additionof this new angiogenic biomarkers patent to our already extensive IP portfolio could enable new methods of monitoring disease progressionandtherapeuticresponse."

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A s biopharmaceutical companies look to cut costs and improve speed- to-market, more of them plan to

outsource R&D and clinical trials and shift this work overseas to places such as China and India, finds a survey released today by global consulting firm Booz & Company.

This study confirms that outsourcing will become an increasingly important part of the competitive landscape and provides key insights into why and how the outsourcing process will radically evolve in the next two to three years. For example, many biopharmaceutical companies will begin outsourcing formerly core activities such as clinical trial monitoring and protocol development to contract research organizations(CROs).Thisshiftwillbedrivenby new service offerings in real-time data processing and virtual platforms that allow people around the world to securely access

Biotech R&D Outsourcing Set to Increase and Shift Overseasclinicaldatainrealtime.Outsourcingmoreof these critical activities along the entire R&D spectrum will transform the nature of outsourcing relationships.

"Thereisaseachangehappeninginthebiopharmaceuticalindustry,"saysMatthewLe Merle, a partner at Booz & Company. "Companies first started to outsource toCROstogetR&Ddonefasterandcheaper.However, this study clearly signals that biotech executives are now looking for more — more value, more expertise, and more innovation. This means we will see outsourcing relationships evolve from transactional to strategic, which will require newcapabilitiesonbothsides."

In mid-2011, Booz & Company partnered with BayBio to conduct a survey and interviews with 32 executives in biopharmaceutical companies. BayBio is Northern California's bioscience association,

representing more than 450 companies.Key findings of the study, called

Outsourcing in Life Sciences: A Survey ofBayBio Members, include a real growth in overseas demand for technology solutions such as real-time data collaboration and globally accessible data storage, and a general trend of clinical trials shifting to China and India in the next two to three years.

"Thesurveyresultsclearlyshowthatoutsourcing is here to stay," says CharleyBeever,apartneratBooz&Company."Thekey question that biopharma companies must address is which capabilities to outsource and which to maintain in-house. Making strategic, coherent decisions about what work is outsourced and why, and how CRO relationships are structured andmanaged, will help companies build a winning business model that adds value, innovation,andcompetitiveadvantage."

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MARCH 20123 – 4 MarchNYSORA Asia 2012Vietnam Contact Person: Mr Yap Hong YiTel: +65 6778 5620Fax: +65 6778 1372Email: [email protected]: http://www.NYSORAAsia.com/

4 – 7 MarchEMBO Symposium. Amoebiasis 2012Khajuraho, India Tel: +91 0 1147165500 Fax: +91 0 11 47165550Email: [email protected]: www.indebo.com

8 – 10 MarchInternational Conference on Biologically Active Molecules 2012 (ICBAM 2012) Dindigul, Tamil Nadu, IndiaContact Person: Dr.N.S.Nagarajan, Director ICBAM 2012Email: [email protected]: +91 9442029933URL: www.icbam2012.com

8 – 11 March20th Annual Meeting of the Asian Society for Cardiothoracic SurgeryBali, IndonesiaContact Person: Latifa HernisaTel: +1162 21566 5993Fax: +1162 21 566 5993Email: [email protected]: www.ascvtsbali2012.org/

12 – 13 March2nd Annual International Conference on Advances in Biotechnology (BIOTECH 2012)Bangkok, Thailand E-mail: [email protected]: www.advbiotech.org

13 – 15 MarchInnovations in Healthcare Management and InformaticsBangkok, ThailandContact: Samuel SeahTel: +65 6722 9388Email: [email protected]: http://www.healthcareinformaticsasia.com/Event.aspx?id=567330

19 – 22 March1st World Congress on Healthy Ageing 2012 Kuala Lumpur, Malaysia Tel: +11 3 2070 5600Email: [email protected]: www.healthyageingcongress.com/

19 – 22 March5th annual BioPharma Asia Convention 2012Marina Bay Sands, SingaporeContact: Valerie LimTel: +65 6322 2766Email: [email protected]: http://www.terrapinn.com/exhibition/biopharma-asi

19 – 22 MarchBiologic Manufacturing World Asia 2012 Marina Bay Sands, Singapore Contact: Valerie Lim Tel: +65 6322 2766 Email: [email protected] URL: http://www.terrapinn.com/conference/biologic-manufacturing-world-asia/

19 – 22 MarchPharma Trials World Asia 2012Marina Bay Sands, SingaporeContact: Valerie LimTel: +65 6322 2766Email: [email protected]: http://www.terrapinn.com/conference/pharma-trials-world-asia/

20 – 24 March15th World Conference on Tobacco or Health SingaporeContact Person: Lysha LimURL: http://wctoh2012.org/

2 – 4 AprilInternational Conference on Food Science and Nutrition 2012 Kota Kinabalu, Sabah, MalaysiaTel: +6088 320000Fax: +6088 320259E-mail: [email protected]: www.ums.edu.my/conferences/ICFSN

12 – 15 April4th Spring Meeting of the International Society for Dermatologic Surgery (ISDS)Tel: +1149 6151 951 8892Fax: +1149 6151 951 8893URL: www.isdsworld.com/en/upcoming-congresses

13 – 15 AprilAsian Oncology Summit 2012Singapore, SingaporeContact Person: Jason LengTel: +65 6349 0201Fax: +65 6733 1817E-mail: [email protected]: http://www.asianoncologysummit.com

April 2012 13 – 16 April27th Asia Pacific Academy of Ophthalmology CongressBusan, South Korea Email: [email protected]: http://www.apaobusan2012.com/

20 – 21 AprilOrganisation for Oncology and Translational Research 8th Annual ConferenceKyoto, Japan Tel: +11 81 75 761 5717Email: [email protected]: www.ootr-institute.org/conference/8th/

20 – 22 AprilThe 9th Meeting of Asian Society for Neuro-Oncology (ASNO2012) Taipei, TaiwanContact Person: Xiao FurenTel: +886 2 2312 3456 ext 63110E-mail: [email protected]: http://www.asno2012.org/

20 – 22 AprilASMMIRT 2012 Sydney, New South Wales, AustraliaContact Person: Lili LinTel: +3 9419 3336Fax: +3 9416 0783Email: [email protected]: www.air.asn.au/asmmirt2012

25 – 26 April2nd Annual Biomarkers in Diagnostics & Therapeutics 2012 SingaporeContact: Ms. Stella AngTel: +65 6853 9156Email: [email protected]: http://www.eventprotocol.com/bdt2012.php

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6 – 8 July1st Asia Pacific Clinical Epidemiology and Evidence Based Medicine Conference Kuala Lumpur, Federal Territory, MalaysiaContact Person: Miss Devi PeramalahTel: +603 7967 3793/3797Fax: +603 7967 4975Email: [email protected]: http://apceebm.um.edu.my

23 – 24 July2012 International Conference on Biological and Life Sciences (ICBLS 2012)SingaporeEmail: [email protected] URL: http://www.icbls.org/

4 – 6 MayWorld Congress on Biotechnology Hyderabad, Andhra Pradesh, IndiaContact Person: Hari KrishnanURL: www.brightice.org

5 – 6 May2012 2nd International Conference on Chemistry and Chemical Process (ICCCP 2012) Kuala Lumpur, MalaysiaE-mail: [email protected]: www.icccp.org/

7 – 9 May7th International Conference on Rapid Response Systems and Medical Emergency Teams Sydney, New South Wales, AustraliaContact Person: Sarah DixonTel: + 61 2 9419 4889Fax: + 61 2 8078 6689URL: http://www.rapidresponsesystems.org/

9 – 11 MayThe 4th International Exhibition on BioPharma, Biotechnology & Equipment 2012Shenzhen, P.R. ChinaContact Person: Ms. Mavis WuTel: +852 2827 6766Fax: +852 2827 6870Email: [email protected] URL: www.coastal.com.hk/biotech

MAY 201216 – 19 May7th World Congress for Neurorehabilitation (WCNR) Melbourne, Victoria, AustraliaTel: +612 9954 4400 Fax: +612 9954 0666URL: www.dcconferences.com.au/wcnr2012

19 – 20 May1st USIM International Conference on Medicine and Health Kuala Lumpur, MalaysiaContact Person: NazefahURL: http://www.icmh2012.usim.edu.my/website

24 – 25 MayNational Medicines Symposium Sydney, NSW, AustraliaContact Person: Maarinke Van Der MeulenTel: +07 3848 2100Fax: +61 7 3848 2133URL: http://www.nps.org.au/nms2012

24 – 27 May19th WONCA Asia Pacific Regional Conference Jeju, Korea (South)Contact Person: Junhee MoonTel: +82 2 566 6033Fax: +82 2 566 6087E-mail: [email protected]: http://www.woncaap2012.org

27 – 28 May17th National Conference on Medical and Health Sciences Kota Bharu, Kelantan, MalaysiaContact person: Abdul Hakim Abdul BasirTel: 09 767 5751Fax: 09 764 2026Email : [email protected]: http://www.dental.usm.my/ncmhs/

27 – 29 MayISPE Singapore ConferenceSingaporeContact: Ms Rachel LowTel: +65 6780 4671Email: [email protected]: http://www.interphexasia.com/ISPEsingaporeconference/

28 – 29 MayInterphex Asia 2012SingaporeContact: Ms Rachel LowTel: +65 6780 4671Email: [email protected]: http://www.interphexasia.com/home/

2 – 3 JuneAdvances in Medicine 2012 Hong Kong, Hong Kong, Hong KongContact name: Richie HeungTel: +852 2632 3127 Fax: +852 2645 1699URL: www.mect.cuhk.edu.hk/AIM2012

5 – 8 JuneRoyal College of Obstetricians and Gynaecologists 10th International Scientific Meeting, RCOG 2012 Kuching, Sarawak, MalaysiaTel: +603 6201 1858Fax: +603 6201 1850Email: [email protected]: www.rcog2012.com

JUNE 20129 -11 June The 3rd International Biotechnology and Biodiversity Conference and Exhibition (BIOJOHOR 2012)Johor Bahru, Johor MalaysiaTel: +607 520 7810 Fax: +607 520 7811/ 3822 Email: [email protected]: http://www.biojohor.my/2012/index.html

30 June – 1 JulyThe 9th International Conference with the Global Network of WHO Kobe, JapanContact Person: Dr. Aiko YamamotoTel: +81 6 6372 3051 Fax: +81 6 6376 2362URL: http://who2012.umin.jp/

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