Banking Human Neural Stem Cells for Clinical Applications. Lisa Fox Director, Cell Process Development & Operations

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1 Banking Human Neural Stem Cells for Clinical Applications Lisa Fox Director, Cell Process Development & Operations 7 th Annual Somatic Cell Therapy Symposium September 27, 2007

2 Stem Cells Inc. Approach for Tissue-Derived Stem Cell Discovery Liver Pancreas Brain Cell Suspension Stained With mabs Proprietary mabs Culture Assays Stem Cell Purification In Vivo Assays Proprietary composition Non-stem cells StemCells, Inc. 2007

3 HuCNS-SC Are Identifiable, Expandable and Bankable Proprietary Expansion Process Different Cell Lines Purify Neural SC population 2.3% 95% All other cells in the brain Expand CELL NUMBER (LOG) DAYS

4 Human Neural Stem Cells in a Bottle Neurons Lysosomal Storage Diseases Spinal Cord Injury Stroke Traumatic Brain Injury Down s Syndrome Alzheimer s Focal Epilepsy ALS Parkinson s Huntington s Astrocytes Lysosomal Storage Diseases Oligodendrocytes Cerebral Palsy Spinal Cord Injury Multiple Sclerosis Stroke Lysosomal Storage Diseases

5 Discovery and Pre-Clinical Development Prospective isolation of human neural stem cells Purity = Reproducibility Characterization of expanded and banked human neural stem cells Preclinical mouse studies Lysosomal storage disorder: enzyme delivery and neuroprotection Spinal cord injury: engraftment, functional recovery, differentiation

6 Characteristics of HuCNS-SC Migrate and engraft reproducibly and robustly Differentiate into the 3 main cell types of CNS cells: NEURONS ASTROCYTES OLIGODENDROCYTES Maintain expected biological activity of the specialized cells (production of enzymes, myelin formation, neurotransmitters, etc) No untoward effects in the CNS (normal karyotype; no tumors in vivo) StemCells, Inc. 2005

7 In-vivo Biological Properties Must Demonstrate: Engraftment and activity Differentiation into the right mature cells that respond naturally to signals in the environment No differentiation into unwanted cells Must Evaluate: Dose and route of administration Compatibility with delivery device Shelf-life of final formulation StemCells, Inc. 2005

8 Safety Toxicology Tumorigenicity analyses No tumors observed (Points to Consider) No tumors observed in CNS Systemic Distribution No abnormal cell growth in other organs examined (lung, spleen, liver, heart, etc) Toxicology Study Transplant cells into brain Monitor physical parameters Monitor behavior No untoward effects Abnormal behavior Tumors

9 Extensive Engraftment, Site Specific Proliferation and Migration of hcns-sc 47 week post transplant Human Specific SC121 mab staining (brown) Transplant Proliferating at neurogeneic site SVZ Migrating as chain of neuroblasts RMS BrdU/Human Nuclei b-tubulin III/ Human nuclei

10 Batten Disease Pre-clinical model Transgenic knockout mouse model (S. Hoffman UT, SMS) recapitulates human condition Deletion of the PPT1 gene loss of palmitoyl protein thioesterase activity Accumulation of autofluorescent storage material Progressive neuronal cell loss Seizures and early death Back-crossed to NOD-Scid mouse StemCells, Inc

11 Migration and differentiation of transplanted hcns-sc progeny in PPT1 KO brains. Immature neural cells Cortical pial Neurons Astrocytes Oligodendrocytes

12 Challenges Using Animal Models of Human Disease Scale Non-comparable anatomy Difficulty performing species specific analyses Xenogeneic rejection despite immunosuppression Immunodeficient model retains human cells long term How applicable is this to the human situation? Few animal models accurately recapitulate the human disease despite widespread use

13 Challenges of Banking of Primary Cell Lines Process Asepsis Feasibility Raw Materials Donor tissue Biologic source Antibodies Cytokines Dissociation enzymes

14 Process Challenges Asepsis Scale / Closing system Open processing steps Primary tissue dissociation Fluorescence activated cell sorting Culture duration Staged release of final product Rapid release Full release

15 Process Challenges, con t Feasibility Containment Processing donor tissue prior to full eligibility determination Jet in Air cell sorting Scale Adequate to cover clinical requirements Schedule Culture Duration Testing and characterization window Shipping logistics

16 Raw Material Challenges - Tissue Donor tissue Donor screening and eligibility determination Non-immortalized cell source Must establish lot to lot comparability

17 Raw Material Challenges - Biologics Biologic Sourced Materials Monoclonal antibodies Antisera Cytokines Fluorochromes Dissociation Enzymes

18 Acknowledgement Kings College Jonathan Cooper Stanford University William Mobley Irving Weissman University of Texas Southwestern Medical School Sandra Hofmann Reeve-UC Irvine Aileen Anderson Brian Cummings