Induced Pluripotent Stemcells: A P4 Perspective as presented by Emile Nuwaysir, PhD; COO and Vice President, Cellular Dynamics International

1. Induced Pluripotent Stem Cells: The Personal Stem Cell Emile F. Nuwaysir, Ph.D. Chief Operating Officer Cellular Dynamics International

2. Induced Pluripotent Stem Cells Introduce four reprogramming factors Blood draw or skin biopsy Induced pluripotent stem (iPS) Cells iPS cells are not the same as embryonic stem cells, and are NOT derived from fertilized embryos! iPS cells are derived from adult tissue sample (skin or blood)

3. Induced Pluripotent Stem CellsTwo Critical Characteristics What’s so special about iPS cells? Replicate Indefinitely Make all 208 cell types in human body Induced Pluripotent Stem (iPS) Cells

4. Reprogramming: Science Magazine’s 2008 Breakthrough of the Year

7. Ethnic diversity

8. Hypo- or hyper-responders

9. “In vitro clinical trials”

10. Any genotype as background for molecular research and medicine… a first for humanity

11. Cardiomyocytes

12. Hepatocytes

13. Neurons

14. Endothelial cells

15. Any cell type for research and medicine… a first for humanityDifferentiate into all 208 cell types in the human body

17. Toxicology (safety)

18. Pharmacology (efficacy)

19. Panels (in vitro Clinical Trials™)

20. Ethnic diversity

21. Genetic predisposition

22. Disease models

23. Genetic

24. Engineered

25. Induced

26. Retrospective analysis of clinical trials

27. Personalized Medicine

28. Autologous cell TherapySource: CDI Analysis

30. Individual drug toxicology (safety)

31. Individual drug pharmacology (efficacy)

32. Disease incidence or progression in target tissue

33. Preventive

34. Adverse drug reactions or disease

35. Personalized

36. 100% personal to you…

37. Participatory

38. Individual donates sample

39. Multiple samples because time-course may be informative

41. Pipeline is applicable across all human tissue types

44. Population scale iPS cell banks (e.g. research cohort, country, etc.)

46. Human cardiomyocytes from iPS cells

47. 99% pure, cryopreserved, ready to use.

48. Complete solution; cells, media, protocols

49. Available in virtually unlimited quantities

50. Express all function cardiac markers, including critical ion channels

52. iCell Cardiomyocytes In Vitro Human Electrophysiology Cells Display Normal Ionic Currents Cells Display Normal Action Potentials Action Potential Examples Atrial-like Nodal-like Ventricular-like Control Tracings 30nM E4031 Control Drug Responses 3 nM Nifedipine 10 nM Nifedipine Control 100 ms

53. iCell CardiomyocyteSingle Cell Pharmacology AP train Selected APs EADs

54. Testimonials “Had stem cell-derived heart tissue been available… the company could have pulled the plug early, saving two years of work and millions of dollars...” - Kyle Kolaja, global head of predictive toxicology screens and emerging technologies for Roche. Sep 23, 2010, BloomBergBusinessWeek, “Stem Cells Test Drugs in a Dish, May Save Companies Millions” Roche "This is a game changer. It's going to dramatically change biology and drug development.” - Sandra Engle, senior principal scientist at Pfizer. Jan 22, 2010, MIT Technology Review “Human heart cells that beat in a dish have never before been available in large quantity and consistent quality. Because they are functional human cells, they may provide a more accurate way of testing drugs than using rodents or the preserved human cells that have been available for lab research in the past,” - Jason Gardner, vice president and head of the stem cell drug performance unit at GlaxoSmithKline. Sep 23, 2010, BloomBergBusinessWeek, “Stem Cells Test Drugs in a Dish, May Save Companies Millions” Pfizer GlaxoSmithKline

56. Human neurons from iPS cells

57. >95% pure, cryopreserved, ready to use.

58. Complete solution; cells, media, protocols

59. Express key molecular and functional attributesTypical morphology observed early after differentiation (left). Three weeks later, the neurons are more organized into extensive networks (below). TH/Hoechst Glutamate/Hoechst GABA/Hoechst

61. Human endothelial cells from iPS cells

62. >95% pure, cryopreserved, ready to use.

63. Full product solution; cells, media, protocols

64. Express key molecular and functional attributesZO-1-488 (green); Hoechst (blue) Inflammation Angiogenesis Stimulated Unstimulated

66. Human hepatocytes cells from iPS cells

67. >95% pure, cryopreserved, ready to use.

68. Full product solution; cells, media, protocols

69. Express key molecular and functional attributes

71. iCell Hepatocyte CharacterizationFunction Lipid Storage Glycogen Storage Albumin Production 3x induction PAS Oil Red 2x induction 60% of iCell Hepatocytes store lipid (BODIPY assay) CYP3A4 Protein Expression CYP3A4 Activity

73. Completed Genome-Wide Association Study - identified SNPs for Left Ventricular Hypertrophy (LVH) and other important phenotypes.

74. Novel approach- use iPSC-derived ventricular cardiomyocytes to study the molecular changes in the development of LVH.

75. First human cardiomyocyte-based cellular model of LVH.

77. Produce 50M cryopreserved ventricular cardiomyocytes from each iPSC line.

78. Experimentally induce the LVH phenotype in culture.

79. Perform RNA expression analysis on genes and that are ‘transcriptional surrogates’ for LVH.

80. PeformeQTL analysis.

81. Determine the role of HyperGen SNPs in LVH epidemiology.

82. Provides a promising approach to test GWA results and common DNA variation in the context of a cell-based system by directly assaying molecular function and effects.

85. Maturity (make terminal cells more mature)

86. Disease phenotypes (derive more disease models)

87. Organotypic biology (2D vs. 3D)

88. Mutlicellular systems (Systems Biology) to “reconstitute” the organ