Doç. Dr. UTKU ATEŞ EUTF HİSTOLOJİ& EMBRİYOLOJİ ABD 1 … · • Professor Department of...
Transcript of Doç. Dr. UTKU ATEŞ EUTF HİSTOLOJİ& EMBRİYOLOJİ ABD 1 … · • Professor Department of...
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Doç. Dr. UTKU ATEŞEUTF HİSTOLOJİ & EMBRİYOLOJİ
ABDEUTF KORDON KANI BANKASI
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Rudolph Virchow:"Omnis cellula e cellula"...
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1900’lu yılların başlarında Avrupalı araştırmacılar çeşitli kan hücrelerinin hepsinin tek bir hücreden KÖKenlendiğini
fark ettiler.
1958 yılında Dr. Min Chueh Chang IVF uygulamasınıtavşanlarda tartışmasız olarak demonstre etti.
1960 embriyonal karsinoma hücreleri bir çeşit anaçhücredir
1963 Ernest McCulloch ve James Till; Fare kemik iliği hücrelerinin txp sonrası kendi kendilerini yenileme
kapasitelerinin olduğunu kantitatif olarak tanımladılar.
1968’de ilk insan yumurtası in vitro olarak fertilize edildi.5
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1978: İlk IVF bebeği olan Louise Brown, İngiltere’de doğdu (25 Temmuz).
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1981:Martin Evans ve Kaufman fare blastosistlerin iç hücre kitlelerinden EKH’leri elde ettiler. İn vitro ortamda pluripotent fare EKH’lerini çoğaltmak için, gerekli kültür şartlarını tanımladılar.
Nature; 292(9);154Nature; 292(9);154--156;1981156;1981. . 7
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1995-96:
Rhesus maymunlarından ve maromosetlerden primat EKH’leri elde
edildi ve bu hücreler, in vitro ortamda yetiştirildi
Thomson J. A., et al.; PNAS USA;92:7844-7848;1995).
James A Thomson
• Professor Department of Anatomy Genome Center of Wisconsin
• Director, Regenerative Biology, Morgridge Inst. for Research8
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1998 yılının en önemli olayı, James Thomson ve ekibinin, IVF laboratuvarında dondurulmuş ya da taze 36 tane embriyondan 5 adet insan embriyonik kök hücre serisi ürettiklerini rapor etmesiydi (
Science, 282 (5391):1145, 1998).
Bu olaydan sonra embriyonik kök hücre araştırmaları ve beraberinde tartışmalar tüm dünyada hız kazandı. 9
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• Science Dergisi 2003 yılının en önemli 10 tıp olayıarasında “Kök Hücreler” ile ilgili gelişmeleri göstermiştir. 10
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FY 2002FY 2002––FY 2010 (Dollars in millions)FY 2010 (Dollars in millions) NIH Stem cell FundingNIH Stem cell Funding
Human Stem CellsHuman Stem Cells NonNon--Human Stem CellsHuman Stem Cells
YearYear EmbryonicEmbryonicNonNon--
EmbryonicEmbryonicEmbryonicEmbryonic
NonNon--EmbryonicEmbryonic
20022002 $10.1$10.1 $170.9$170.9 $71.5$71.5 $134.1$134.1
2003 2003 $20.3$20.3 $190.7$190.7 $113.5$113.5 $192.1$192.1
2004 2004 $24.3$24.3 $203.2$203.2 $89.3$89.3 $235.7$235.7
2005 2005 $39.6$39.6 $199.4$199.4 $97.1$97.1 $273.2$273.2
2006 2006 $37.8$37.8 $206.1$206.1 $110.4$110.4 $288.7$288.7
2007 2007 $42.1$42.1 $203.5$203.5 $105.9$105.9 $305.9$305.9
2008 2008 $88.1$88.1 $297.2$297.2 $149.7$149.7 $497.4$497.4
2009 (Non2009 (Non--ARRA) ARRA)
$119.9$119.9 $339.3$339.3 $148.1$148.1 $550.2$550.2
2009 (ARRA) 2009 (ARRA) $22.7$22.7 $57.9$57.9 $29.1$29.1 $88.1$88.1
2010 (Non2010 (Non--ARRA) ARRA)
$125.5$125.5 $340.8$340.8 $175.3$175.3 $569.6$569.6
2010 (ARRA) 2010 (ARRA) $39.7$39.7 $73.6$73.6 $19.6$19.6 $74.2$74.211
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1.Advanced genomics and its applications for health (€ 1100m)
1.1 Fundamental knowledge and basic tools for functional genomicsin all organisms
1.2 Applications of knowledge and technologies in the field of genomics and biotechnology for health
(includes SC research and regenerative medicine)
2.1 Application oriented genomic approaches to medical knowledge and technologies
2.2 Combating cancer
2.3 Confronting the major communicable diseaseslinked to poverty
2. Combating major diseases (1155m)
Life Sciences and Biotechnology for Health
~ €2.4 billion in 6th Framework programme
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REGENERATIVE MEDICINE
Cord Blood
Mesenchymal SC
hESC
Tissue repair& regeneration
skinbrain spine
islets
Soft tissue
hematopoietic SC
heart
muscle
Transplants
Mechanisms
Banking
ALLOSTEMImmunotherapeutic strategies
to treat haematological and neoplastic diseases with
optimised allogeneic stem cell transplantation (IP, € 8m)
www.allostem.org
GENOSTEM:Adult mesenchymal stem cell
engineering for connective tissue disorders (IP, € 8.6m)
www.genostem.org
EUROSTEMCELL:European Consortium for Stem
Cell Research (IP, € 12m)www.eurostemcell.org
FUNGENES:Functional Genomics in Embryonic Stem Cells
(IP, € 12m)www.fungenes.org
CRYSTAL:CRYo-banking of Stem cells for human Therapeutic AppLication
(STREP, € 2.4m)
BETACELLTHERAPY:Beta Cell Programming for
Treatment of Diabetes (IP, €12m)
SC&CR:Application and process
optimisation of human stem cells for myocardium repair
(STREP, € 2m)
RESCUE:From stem cell technology to
functional restoration after spinal cord injury (STREP, €
2.7m)
OSTEOCORD:Bone from blood:
mesenchymal stem cells from umbilical cord blood
(STREP, € 2.5m)
THERCORD:Development and preclinical testing of cord blood-derived
cell therapy products (STREP, € 1.8m)
STEMSTROKE:Towards a stem cell therapy for stroke (STREP, € 2.5m)
STEM-HD:Embryonic stem cells for
therapy and exploration of mechanisms in Huntington Disease (STREP, € 2.5m)
Ulcer Therapy:Gene Transfer in Skin
Equivalents and Stem Cells for Chronic Ulcer Repair and
Tissue Regeneration (STREP, € 2.5m)
Stem Cell research in FP6in the “New Therapies” area
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• Budget increase gradually from €600m/yearP to > €1
billion/year
• Collaborative Research (Cooperation)
� Continuity in themes & instruments, longer duration� Funding levels (50%) to be raised in specific cases (+25%) � Simplification of procedures (flat rates; collective responsibility)
• European Research Council (Ideas)
• Joint Technology Initiative for Innovative Medicines
Health research in FP 7
The European Commission currently manages about 5% of total public spending in R&D in the European Union
The European Union currently invests about 1.9% of GDP in research
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NEDEN,
KÖK HÜCRELER
BU KADAR ÖNEMLİ? 16
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BU HÜCRELERLE HASTALIKLAR NASIL TEDAVİ EDİLEBİLİR?
• REJENERATİF TIP
• REPARATİF TIP 17
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BİR HEDEF DOKU VEYA ORGAN HASARINDA;
O organın işlevlerini eski haline getirmeye yetecek sayı ve kalitede izole edilmiş ve özellikleri belirlenmiş olan sağlıklıhücrelerin nakledilmesiyle tedavi sağlanabilir. 18
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KKÖÖK HK HÜÜCRELER, CRELER,
HHÜÜCRE TABANLI TEDAVCRE TABANLI TEDAVİİ PROTOKOLLERPROTOKOLLERİİNNİİN N VAZGEVAZGEÇÇİİLMEZ UNSURLARIDIRLMEZ UNSURLARIDIROO
NEDEN????NEDEN???? 19
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ÇÜÇÜNKNKÜÜ, ,
BU HBU HÜÜCRELERCRELERİİ VVÜÜCUDUMUZUN DCUDUMUZUN DİĞİĞER SOMATER SOMATİİK HK HÜÜCRELERCRELERİİNDEN NDEN
FARKLI KILAN FARKLI KILAN İİKKİİ ÖÖNEMLNEMLİİ ÖÖZELLZELLİĞİİĞİ VAR;VAR;
�� ÇÇOOĞĞALABALABİİLMELME--BBÖÖLLÜÜNEBNEBİİLME,( SELFLME,( SELF--RENEWAL)RENEWAL)
�� PLASTPLASTİİSSİİTE (DTE (DİİFFERENSFFERENSİİASYONASYON--TRANSDTRANSDİİFFERENSFFERENSİİASYON).ASYON). 20
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1 - Simetrik kök hücre bölünmesi;
2 - Asimetrik kök hücre bölünmesi;
3 - Progenitor bölünme;
4 - Terminal farklılaşma
1,2 => (SELF-RENEWAL)
2,3,4 => (FARKLILAŞMA)
Kök hücre bölünme ve farklılaşma tablosuA – kök hücre; B - progenitor hücre; C – farklılaşmış hücre;
Kök hücre bölünme ve farklılaşma tablosu
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• SC’in diğer hücrelerle etkileşimi
• Adhezyon molekülleri, • EC matriks bileşenleri; • Büyüme faktörleri;• Sitokinler;• Mikroçevre
FARKLILAFARKLILAŞŞMA MA
ÇÇOOĞĞALMAALMA
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YÖNLENDİRİLMİŞFARKLILAŞMA
ÖZEL KÜLTÜR KOŞULLARI
KÖK HÜCRE
OLGUN HÜCRE (SİNİR)
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A. TRANSDETERMİNASYON:
Manohar R.et al. Transdetermination: A New Trend in Cellular Reprogramming Molecular Therapy; vol. 17 no. 6 june 2009
*(Ngn3); endocrine transcription factor neurogenin 3
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B. TRANSDİFERANSİYASYON:
TRANSDİFERANSİYASYON
DİF
ER
AN
SİY
AS
YO
N
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C. DEDC. DEDİİFERANSFERANSİİYASYON:YASYON:
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D. HÜCRE FÜZYONU:
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Farklı kaynaklardan elde edilen kök hücrelerin farklılaşma kapasiteleri arasında da fark vardır:
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1.Totipotent
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2.Pluripotent Kök Hücre:
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İndüklenmiş Pluripotent Kök Hücre:
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3. Multipotent KH:
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4. Oligopotent KH:
VASKÜLER KÖK HÜCRE
ENDOTEL HÜCRESİ
DÜZ KAS HÜCRESİ37
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5. Unipotent KH:
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a) Embriyonik Kök Hücreler
b) Embriyonik Olmayan Kök Hücreler
Bu gBu güüne kadar, kne kadar, köök hk hüücrelerin kaynaklarcrelerin kaynaklarıına yna yöönelik farklnelik farklııssıınnııflamalar tanflamalar tanıımlanmmlanmışışttıır. r.
Colonies of embryonic stem cells - University Of Wisconsin-Madison39
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I – Hematopoietik Kök Hücreler
a – Kemik İliği Kök Hücrelerib – Periferik Kan Kök hücreleric – Göbek Kordon Kanı Kök Hücreleri
II – Stromal (Mezenkimal) Kök Hücreler
III – Organlarda yerleşik diğer erişkin kök hücreleri
A) Embriyonik Kök Hücreler
B) Embriyonik Olmayan Kök Hücreler (Erişkin kök hücreleri)
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Embriyonik geliEmbriyonik gelişşimin erken evrelerinde embriyondan elde imin erken evrelerinde embriyondan elde edilen kedilen köök hk hüücrelerdir. (Ycrelerdir. (YÜÜT; IVFT; IVF--Klonlama)Klonlama)
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Alt kültürleme (pasaj).
30 hc.���� milyonlarca hc.
Tek ���� hc.serisi ����klonal
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EMBRİYONİK OLMAYAN KÖK HÜCRELER
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HEMATOPOİETİK KÖK HÜCRELER (HSC)
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HEMATOPOİETİK KÖK HÜCRE -
HEMATOPOEZ
0 2 4 6 8 91 3
Karaciğer-Dalak
Kemik İliği
5 7 AY
Kan Yapım Yeri
Yolk Kesesi
AGM BÖLGESİ
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HKH-HEMATOPOEZ
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HEMATOPOEZ-HKH
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HSC + FİBRİLER KOLLAGEN
HEMATOPOİETİK KÖK HÜCRE;
KEMİKİ İLİĞİ 1/10.000PERİFERİK KAN !/100.000KORDON KANI 1/100
SELF RENEWAL
APOPTOZİS
PLURİPOTENT
MOBİLİZASYON VE HOMİNG
PLASTİSİTE
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KEMİKİLİĞİ MİKROÇEVRESİ
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Mezey E et al: PNAS 2003, 100:1364-136960
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Kocher et al; Nature Medicine 7, 430 - 436 (2001) 61
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Rejenerasyondaki hepatik nodüllerin immunfloresans boyaması (FAH-/- mice.)
Lagasse E et al; Nature Medicine 2000, 6:1229-123462
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Iannus et al; J Clin Invest 2003, 111:843-85063
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II-STROMAL (MEZENKİMAL) KÖK HÜCRE
M F Pittenger et al. Science 1999;284:143-147
• KEMİK İLİĞİ• GÖBEK
KORDONU• PERİOSTEUM• ADİPOZ DOKU• DERMİS• KAS• DAMARLAR• PERİODONTAL
LİGAMENT
MSC
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III-ORGANLARDA YERLEŞİK KÖK HÜCRELER
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