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Embryomics: Commercial Opportunities in the Increasingly Complex Biology of Pluripotency Case Western Reserve University

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Embryomics: Commercial Opportunities in the Increasingly Complex Biology of Pluripotency Case Western Reserve University Embryomics: Commercial Opportunities in the Increasingly Complex Biology of Pluripotency Case Western Reserve University July 16, 2013 Forward Looking Statements The matters discussed in this presentation include forward looking statements which are subject to various risks, uncertainties, and other factors that could cause actual results to differ materially from the results anticipated. Such risks and uncertainties include but are not limited to the success of BioTime in developing new stem cell products and technologies; results of clinical trials of BioTime products; the ability of BioTime and its licensees to obtain additional FDA and foreign regulatory approval to market BioTime products; competition from products manufactured and sold or being developed by other companies; the price of and demand for BioTime products; and the ability of BioTime to raise the capital needed to finance its current and planned operations. Any statements that are not historical fact (including, but not limited to statements that contain words such as "will," "believes," "plans," "anticipates," "expects," "estimates") should also be considered to be forward-looking statements. Forward-looking statements involve risks and uncertainties, including, without limitation, risks inherent in the development and/or commercialization of potential products, uncertainty in the results of clinical trials or regulatory approvals, need and ability to obtain future capital, and maintenance of intellectual property rights. As actual results may differ materially from the results anticipated in these forward-looking statements they should be evaluated together with the many uncertainties that affect the business of BioTime and its subsidiaries, particularly those mentioned in the cautionary statements found in BioTime's Securities and Exchange Commission filings. BioTime disclaims any intent or obligation to update these forward-looking statements. 2 The Opportunity in Pluripotency • Scalable source of all human cell types • Source of embryonic progenitors 3 Challenges of Pluripotency • Scalable & reproducible product • Purity and identity of cells • A formulation optimizing viability & immobilization of engraftment • Strategies for near-term commercialization 4 Contrasted Scale-Up Strategies Purification of Problem of impurities Differentiation ES Cells desired cell type >200-fold diversity Scalable, monoclonally purified progenitors 5 Solving the Challenge of Reproducibility Embryoid Body Activin A FGF2 Wnt 3A Pluripotent Stem Cells 1,000 + Complexity of Cell Types 6 The Opportunity of hES Cell Scalability TRF Length Population Doublings (kbp) Germ-Line & Pluripotent SCs 22 34 43 55 65 72 82 90 15 (Telomerase +) Somatic 10 (Telomerase -) 5 Decreasing Telomere Length with Age Length with Telomere Decreasing Neonatal Hayflick Limit Age 7 Solving the Technical Hurdles 8 Solving the Technical Hurdles 9 Solving the Technical Hurdles 10 Solving the Technical Hurdles 11 Manufacturing Technology 2.0 Human embryonic progenitor (hEP) cell lines: > 200 diverse cell types isolated Kidney Diversity Smooth Muscle Precise identity Skeletal Muscle Purity Scalability Blood/Brain Patents pending Barrier 12 Purity Purified Embryonic Progenitor Line E68 CNTN6 CNTN6 Isotype Ab Reg Med 3(3):287 (2008) Direct Scalability Selection Criteria of Scalability in Rollers hEP Cell Line Scalability Regen Med 2012 Jul;7(4):481-501 4D20.8 Scalability Regen Med 2012 Jul;7(4):481-501 4D20.8 Scalability Fate Space Screening TGFbs Chondrogenesis FGFs Angiogenesis BMPs Osteogenesis RA Neurogenesis WNTs Myogenesis >100 Scalable Array of Diverse Approx 4,000 Clonal hEP Differentiation Gene Expression Lines Conditions Microarrays 18 Fate Space Screening T42 in MM Culture T42 in HyStem Culture 19 Subsidiaries OncoCyte Corporation Cancer diagnostics and anti-cancer therapies Cell Cure Neurosciences Ltd. Neurology OrthoCyte Corporation Orthopedics ReCyte Therapeutics, Inc. Age-related vascular disorders ES Cell International PTE Ltd. Research products LifeMap Sciences, Inc. Online databases Asterias Biotherapeutics Contribution of Geron and BioTime Assets 20 Osteochondral Differentiation Micromas s with TGFb3 COL2A1 qPCR Condition7SMOO32 4D20.8 MSCs SM30 7PEND24 E15 MEL2 SK11 NHAC 21 Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines Osteochondral Differentiation Micromas s with TGFb3 Condition 24 PureStem Line 4D20.8 • HOXB2+ • BARX1+ • LHX8+ • FOXF2- 25 4D20.8 hEP Reproducibility 26 Solving the Manufacturing Challenges Simplified and more reproducible differentiation FGF2 Wnt 3A 1,000 + Complexity of Cell Types 27 Osteochondral Differentiation DPSCs 4D20.8 28 Precise Identity Foxf1 Lhx8 Barx1 Foxf1 Genes & Dev. 18: 937-951 Lhx8 Science 24:306: 2255-2257 Barx1 Development 136: 637-645 29 Osteochondral Differentiation 30 Osteochondral Differentiation 31 COL2A1 (Cartilage) Induction * * * 32 Regen Med 2012 Jul;7(4):481-501 Diverse Osteochondral hEP Cell Lines Effects of GDF5 PureStem Line E15 NNAT+ Chondrogenic Progenitor E15 Differentiated in GDF5 Positive For: Negative For: NNAT MSX1 HOXA2 FOXF1 HOXB2 FOXF2 PureStem Line SM30 • HOX- • PITX1+ • ZIC2+ • TBX15- 35 PureStem Lines SK11 AND SM30 PITX1 ZIC2 TBX15 SK11 + + + SM30 +PITX1 + ZIC2 TBX15 - Genepaint.org Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines E15 Diversity of Defined Osteochondral Cells 40 Limb Bud Markers We are beginning to understand limb morphogenesis as controlled by site- specific homeobox genes and gradients from morphogen fields. Such lines, if capable of osteochondral differentiation, could have the advantages of homologous use (i.e. not putting mandibular mesenchyme in the knee). First, limb buds are an outgrowth from lateral plate mesoderm (LPM), specifically the somatic mesoderm). Splanchnic Mesoderm Lateral Plate Mesoderm Somatic Mesoderm 41 Limb Bud Markers The nomenclature is as follows: 42 Limb Bud Markers Distal LPM, perhaps from distal HOX genes, is thought to trigger PITX1, a dominant factor for conversion of limb bud to hindlimb. PITX1 is also expressed in mandibular mesenchyme as shown below for PureStem SK11 and SM30. Mandibular Mesenchyme Forelimb Hindlimb Taher L, Collette NM, Murugesh D, Maxwell E, Ovcharenko I, et al. (2011) 43 Limb Bud Markers HOXB6 as a marker of committed limb bud mesenchyme 44 Limb Bud Markers HOXA10 expression in limb mesenchyme 45 Limb Bud Markers NHAC RTPEC ASCs EN1 RA Series SK17 MSCs B16 C4ELSR10 SK31 T14 4SKEL20 46 Limb Bud Markers HOXA13 as a marker of distal limb mesenchyme 47 Limb Bud Markers NHAC C4ELSR10 Xgene EN1 RAD20.19 SK17 B16 SK31 T14 MSCs C4ELS5.1 48 Novel Osteogenic Lines E3 & E72 Embryonic Forelimb Mesenchyme (EFM) E3 E72 49 Novel Osteogenic Lines E72 & E75 Embryonic Forelimb Mesenchyme (EFM) E3 EFM E3 EFM 50 Scalable Human Embryonic Limb Cells Upper Limb E72 Lower Limb RAD20.5 HOXA10+ HOXA10+ HOXD11 HOXB6+ PITX1- PITX1+ 51 Scalable Human Embryonic Limb Cells Iliac MSCs Lower Limb RAD20.5 HOXA10+ HOXA10+ HOXB6+ HOXB6+ PITX1+ PITX1+ CD74+ CD74- 52 PureStem E69 and T42 53 Potential Uses of PureStem E69 & T42 E69 T42 54 Potential Uses of PureStem E69 & T42 CYP26B1 TTR Basio-Occipital Bone Choroid Plexus Meninges Of 4th Ventricle Of Upper Medulla In situ images from Genepaint.org 55 Storage Strategies Cells can be differentiated and frozen in HyStem 56 LifeMap 57 Orthopedic Applications > 30 distinct osteochondro-progenitors BACK PAIN Degenerative disc disease: Disc repair/regeneration Spinal fusion: Bone induction Back Pain DDD KNEE PAIN: Cartilage injuries, bone defects, osteoarthritis Meniscus, ligament, Tendon Knee Pain Traumatic injury 58 PureStem Manufacturing Technology Human embryonic progenitor (hEP) cell lines: > 200 diverse cell types isolated • Reproducible product Kidney • Purity and identity • Can differ from MSCs in Smooth Muscle Skeletal Muscle being non-hypertrophic • Potential for permanent Blood/Brain engraftment Barrier • Improved survival and translatability with HyStem 59.
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