ArunA BiomedicalSTEMEZ™ hNP1™, hN2™ and iPS-NP1™

Demonstrated use in HTS and HCA assay systems

April 26, 2011

PROPRIETARY AND CONFIDENTIAL

Overview of ArunA’s neural cell products

STEMEZ™ hNP1™ Human Neural Progenitor Cells

Tissue source: Human embryonic stem cells

Culture system: Highly proliferative adherent mono-layer

Feeder system: Feeder and serum free cultures

Differentiation potential: Dopaminergic, cholinergic, glutamatergic, GABA-ergic and astrocytic lineages

STEMEZ™ hN2 Human Neural CellsTissue source: Human embryonic stem cellsCulture system: Adherent mono-layerFeeder system: Feeder and serum free culturesDifferentiation potential: hN2 cells are a mixed population of differentiated neuronal cells

STEMEZ™ iPS-NP1™

Tissue source: Human induced pluripotent stem cells from lung fibroblasts

Culture system: Highly proliferative adherent mono-layer

Feeder system: Feeder and serum free cultures

Differentiation potential: Mixed population of neuronal cells. Further studies are underway.

PROPRIETARY AND CONFIDENTIAL 2

Application of neural cells to HTS ATP assay

Objective:

To test ATP levels in hN2™ , hNP1™ and iPS-NP1™cells using HemoGenix HTS LUMISTEM™ assay kit

Procedure:• 96-well plates were coated with Matrigel (1:200) for ~1 hour at 4°C and washed

twice with PBS.• Cells were plated at 10,000 cells per well and incubated at 37°C for ~48 hrs then

assayed for ATP levels, as directed in the assay kit protocol.

PROPRIETARY AND CONFIDENTIAL 3

Cellular ATP levels in ArunA’s neural cells

• STEMEZ™ hNP1™ and STEMEZ™ iPS-NP1™ STEMEZ neural progenitor cells and hN2™ differentiated neural cells are amenable to HTS ATP analysis using the LUMISTEM assay.

Cell Type Mean [ATP] SD %CV

hNP1™ 1.15 µM 0.03 µM 2.4%

iPS-NP1™ 1.11 µM 0.02 µM 2.1%

hN2™ 0.68 µM 0.01 µM 2.2%

PROPRIETARY AND CONFIDENTIAL 4

Data in collaboration with HemoGenix

STEMEZ™ hNP1™ and hN2™ use in HCA assays

Objective: • Evaluate the applicability of STEMEZ™ hNP1™ and hN2™ cells in HCA

neurite outgrowth assay to assess:– Neurotoxicity (hN2™)– Neuronal differentiation

Procedure and outcomes:• Molecular Devices’ ImageXpress platform and associated neurite outgrowth

scoring modules were used for image acquisition and analysis• Both cell types proved to be effective choices for automated HTS formatted

compound testing

PROPRIETARY AND CONFIDENTIAL 5

Automated measurement of neurite outgrowth in hN2™. Neuritesemerging from accepted cell bodies are traced (light blue, green and purple lines) and quantified while cells without process (red) are not counted

PROPRIETARY AND CONFIDENTIAL 6

Harrill JA, Freudenrich TM, Machacek DW, Stice SL, Mundy WR. (2010) Quantitative assessment of neurite outgrowth in human embryonic stem cell-derived hN2 cells using automated high-content image analysis. Neurotoxicology. 31(3):277-90.

Application of hN2™ neural cells to HCA neurite outgrowth assay

Green, beta III-tubulin (neurites)Blue, Hoechst (nuclei)

Image analysis of neurite outgrowth in hN2™ cells using ImageXpress

Nuclei and neurite traces

PROPRIETARY AND CONFIDENTIAL 7

Data in collaboration with Molecular Devices

Effect of Antimycin A on hN2™ neurite outgrowth

0 µM

1 µM

10 µM

100 µM

PROPRIETARY AND CONFIDENTIAL 8

Compound effects on hN2™ neurite outgrowth

PROPRIETARY AND CONFIDENTIAL 9

Differentiation of hNP1™ for 14 days in 96-well format

PROPRIETARY AND CONFIDENTIAL 10

Application of differentiated hNP1™ cells to HCA neurite outgrowth assay

Nuclei and neurite traces

PROPRIETARY AND CONFIDENTIAL 11

Green, beta III-tubulin (neurites)Blue, Hoechst (nuclei)

Data in collaboration with Molecular Devices

Effect of LIF on neurite outgrowth in 14-day differentiated hNP1™ cells

0 50 100 150 200 250

max process length

mean process length

total outgrowth

Length (µm)

-LIF

+LIF

PROPRIETARY AND CONFIDENTIAL 12

Data in collaboration with Molecular Devices