Personalized progenitor cells

Abstract

Tissue specific progenitor cells generated from pluripotent sources representing endoderm, ectoderm and mesoderm lineages. Progenitor generation can be accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins and peptides obtained from stressing said tissue ex vivo. Ex vivo generated tissue organoids which are utilized as sources of "differentiation factors" for the creation of personalized progenitor cells. For generation of pulmonary progenitor cells, decellularized cadaveric lung is populated with allogeneic stem cells to form pulmonary structures, said structures are exposed to various cellular stressors and conditioned media to differentiate pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerative cells and used to create organoids that are subjected to stress in order to generate differentiating factors.

Description

PERSONALIZED PROGENITOR CELLSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims benefit of U.S. Provisional Patent Application Serial No. 63 / 599,440, filed on November 15, 2023, entitled "Personalized Progenitor Cells", the contents of which are incorporated herein by reference in its entirety.FIELD OF THE INVENTION

[0002] The invention pertains to the field of regenerative medicine, more particularly the invention pertains to generation of progenitor cells from pluripotent stem cells, more particularly the invention pertains to generation, expansion and use of tissue specific progenitor cells from pluripotent stem cells. The invention pertains to the field of regenerative medicine, more particularly the invention pertains to generation of progenitor cells from pluripotent stem cells, more particularly the invention pertains to generation, expansion and use of tissue specific progenitor cells from pluripotent stem cells.BACKGROUN D OF TH E INVENTION

[0003] Stem cell therapeutics has provided a paradigm shift in medical practice, offering for the first time the possibility of curing previously uncurable diseases by creation of new tissues. Administration of allogeneic stem cells has provided some therapeutic signals, however, this modality is limited by the fact that these cells possess a relatively short in vivo viability. In fact, some have demonstrated that allogeneic stem cell therapeutic effects are not the result of cellular effects but instead elicited by apoptotic bodies of the cells. The invention provides means of using autologous cells for therapeutic uses through the generation of dedifferentiated pluripotent stem cells and creation of tissue specific progenitors that can be banked or used therapeutically when needed.SUMMARY OF TH E INVENTION

[0004] A summary is provided below based on numbered aspects of the invention.

[0005] 1. A method of generating tissue specific progenitors from pluripotent stem cells comprising exposing said pluripotent stem cells to factors generated by representative tissue cells.

[0006] 2. The method of aspect 1, wherein said pluripotent stem cells are capable of forming a teratoma in an immune deficient mouse.

[0007] 3. The method of aspect 1, wherein said pluripotent stem cells are generated by parthenogenesis.

[0008] 4. The method of aspect 3, wherein said parthenogenesis is performed by induction of calcium flux in one or more oocytes.

[0009] 5. The method of aspect 4, wherein said calcium flux is induced by administration of a calcium ionophore.

[0010] 6. The method of aspect 5, wherein said calcium ionophore is A23187.

[0011] 7. The method of aspect 1, wherein said pluripotent stem cells are generated by somatic cell nuclear transfer.

[0012] 8. The method of aspect 1, wherein said pluripotent stem cells are generated by induced pluripotency.

[0013] 9. The method of aspect 8, wherein said induced pluripotency is endowed by transfection of a cell with dedifferentiation factors.

[0014] 10. The method of aspect 9, wherein said dedifferentiation factors are pluripotency associated genes.

[0015] 11. The method of aspect 10, wherein said pluripotency associated gene isOCT4.

[0016] 12. The method of aspect 10, wherein said pluripotency associated gene isNANOG.

[0017] 13. The method of aspect 10, wherein said pluripotency associated gene is KLF.

[0018] 14. The method of aspect 10, wherein said pluripotency associated gene isHER2.

[0019] 15. The method of aspect 10, wherein said pluripotency associated gene is SOX-2.

[0020] 16. The method of aspect 10, wherein said pluripotency associated gene isPIM1.

[0021] 17. The method of aspect 10, wherein said pluripotency associated gene is hTERT.

[0022] 18. The method of aspect 8, wherein said pluripotency is induced by administration of a pluripotency factor together with one or more dedifferentiation factors.

[0023] 19. The method of aspect 18, wherein said dedifferentiation factor is conditioned media from an embryonic stem cell.

[0024] 20. The method of aspect 18, wherein said dedifferentiation factor is conditioned media from a pluripotent stem cell.

[0025] 21. The method of aspect 18, wherein said dedifferentiation factor is conditioned media from a pluripotent stem cell that has been exposed to cellular stress.

[0026] 22. The method of aspect 21, wherein said cellular stress is selected from a group of stressors comprising of: a) hypoxia; b) metabolic stress; c) hyperthermia; d) hypothermia; e) hypotonic stress; f) hypertonic stress; and e) proteosome blockade stress.

[0027] 23. The method of aspect 18, wherein said dedifferentiation factor is conditioned media from a mesenchymal stem cell.

[0028] 24. The method of aspect 23, wherein said mesenchymal stem cell expressesCD73.

[0029] 25. The method of aspect 23, wherein said mesenchymal stem cell expressesCD105.

[0030] 26. The method of aspect 23, wherein said mesenchymal stem cell expresses c- met.

[0031] 27. The method of aspect 23, wherein said mesenchymal stem cell expresses leukemia inhibitor factor.

[0032] 28. The method of aspect 23, wherein said mesenchymal stem cell expresses IL-10 receptor.

[0033] 29. The method of aspect 23, wherein said mesenchymal stem cell expressesSTAT3.

[0034] 30. The method of aspect 23, wherein said mesenchymal stem cell expressesFoxP3.

[0035] 31. The method of aspect 23, wherein said mesenchymal stem cell expressesAIRE.

[0036] 32. The method of aspect 23, wherein said mesenchymal stem cell expresses indolamine 2,3 dioxygenase.

[0037] 33. The method of aspect 23, wherein said mesenchymal stem cell expressesCD132.

[0038] 34. The method of aspect 23, wherein said mesenchymal stem cell expresses c- kit.

[0039] 35. The method of aspect 23, wherein said mesenchymal stem cell expressesPD-L1.

[0040] 36. The method of aspect 23, wherein said mesenchymal stem cell expressesTGF-beta.

[0041] 37. The method of aspect 23, wherein said mesenchymal stem cell expresses endoglin.

[0042] 38. The method of aspect 23, wherein said mesenchymal stem cell expresses arginase.

[0043] 39. The method of aspect 23, wherein said mesenchymal stem cell expresses galectin-1.

[0044] 40. The method of aspect 23, wherein said mesenchymal stem cell expresses galecin-3.

[0045] 41. The method of aspect 23, wherein said mesenchymal stem cell expresses galectin-7.

[0046] 42. The method of aspect 23, wherein said mesenchymal stem cell expressesTIMP-1.

[0047] 43. The method of aspect 23, wherein said mesenchymal stem cell expressesTIMP-3.

[0048] 44. The method of aspect 23, wherein said mesenchymal stem cell expresses urokinase plasminogen activator.

[0049] 45. The method of aspect 23, wherein said mesenchymal stem cell expresses protein C.

[0050] 46. The method of aspect 23, wherein said mesenchymal stem cell expresses antithrombin-IIL

[0051] 47. The method of aspect 23, wherein said mesenchymal stem cell expresses soluble HLA-G.

[0052] 48. The method of aspect 23, wherein said mesenchymal stem cell expresses plasminogen.

[0053] 49. The method of aspect 23, wherein said mesenchymal stem cell expresses IL-7 receptor.

[0054] 50. The method of aspect 23, wherein said mesenchymal stem cell expresses interleukin-12 p40 homodimer.

[0055] 51. The method of aspect 23, wherein said mesenchymal stem cell does not express H LA-11.

[0056] 52. The method of aspect 23, wherein said mesenchymal stem cell does not express H LA-DR.

[0057] 53. The method of aspect 23, wherein said mesenchymal stem cell does not express CD14.

[0058] 54. The method of aspect 23, wherein said mesenchymal stem cell does not express CD16.

[0059] 55. The method of aspect 23, wherein said mesenchymal stem cell does not express CD34.

[0060] 56. The method of aspect 23, wherein said mesenchymal stem cell is isolated by expression of CD73 and subsequently expanded.

[0061] 57. The method of aspect 23, wherein said mesenchymal stem cell is isolated by expression of c-met and subsequently expanded.

[0062] 58. The method of aspect 23, wherein said mesenchymal stem cell is isolated by expression of c-kit and subsequently expanded.

[0063] 59. The method of aspect 23, wherein said mesenchymal stem cell is isolated by expression of DAF and subsequently expanded.

[0064] 60. The method of aspect 23, wherein said mesenchymal stem cell is isolated by expression of interleukin-10 receptor and subsequently expanded.

[0065] 61. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from omental tissue.

[0066] 62. The method of aspect 61, wherein said omental tissue is degraded by collagenase and / or trypsin before isolation of said cells.

[0067] 63. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from bone marrow.

[0068] 64. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from cadaveric bone marrow.

[0069] 65. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from dermal tissue.

[0070] 66. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from adipose tissue.

[0071] 67. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from placental tissue.

[0072] 68. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from chorionic placental tissue.

[0073] 69. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from umbilical cord tissue.

[0074] 70. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from subepithelial umbilical cord tissue.

[0075] 71. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from peripheral blood.

[0076] 72. The method of aspects 71, wherein said peripheral blood is obtained after mobilization.

[0077] 73. The method of aspect 72, wherein said mobilization is induced by treatment with an inhibitor of the SCF-1 / CXCR4 interaction.

[0078] 74. The method of aspect 73, wherein said mobilizing agent is a small molecule inhibitor of CXCR4.

[0079] 75. The method of aspect 74, wherein said small molecule inhibitor of CXCR4 isMozibil.

[0080] 76. The method of aspect 72, wherein said mobilizing agent is a small interferingRNA targeting CXCR4.

[0081] 77. The method of aspect 72, wherein said mobilizing agent is a small interferingRNA targeting SDF-1.

[0082] 78. The method of aspect 72, wherein said mobilizing agent is a small hairpinRNA targeting CXCR4.

[0083] 79. The method of aspect 72, wherein said mobilizing agent is a small hairpinRNA targeting SDF-1.

[0084] 80. The method of aspect 72, wherein said mobilizing agent is a small interferingRNA targeting CXCR4.

[0085] 81. The method of aspect 72, wherein said mobilizing agent is a small interferingRNA targeting SDF-1.

[0086] 82. The method of aspect 72, wherein said mobilizing agent is a ribozyme targeting CXCR4.

[0087] 83. The method of aspect 72, wherein said mobilizing agent is a ribozyme targeting SDF-1.

[0088] 84. The method of aspect 72, wherein said mobilizing agent is a hammerhead ribozyme targeting CXCR4.

[0089] 85. The method of aspect 72, wherein said mobilizing agent is a hammerhead ribozyme targeting SDF-1.

[0090] 86. The method of aspect 72, wherein said mobilizing agent is an antisense oligonucleotide molecule targeting CXCR4.

[0091] 87. The method of aspect 72, wherein said mobilizing agent is an antisense oligonucleotide molecule targeting SDF-1.

[0092] 88. The method of aspect 72, wherein said mobilizing agent is cyclosporine

[0093] 89. The method of aspect 72, wherein said mobilizing agent is cyclophosphamide.

[0094] 90. The method of aspect 72, wherein said mobilizing agent is VEGF.

[0095] 91. The method of aspect 72, wherein said mobilizing agent is flt-3 ligand.

[0096] 92. The method of aspect 72, wherein said mobilizing agent is interleukin-11.

[0097] 93. The method of aspect 72, wherein said mobilizing agent is TNF-alpha.

[0098] 94. The method of aspect 72, wherein said mobilizing agent is G-CSF.

[0099] 95. The method of aspect 72, wherein said mobilizing agent is M-CSF.

[0100] 96. The method of aspect 72, wherein said mobilizing agent is GM-CSF.

[0101] 97. The method of aspect 72, wherein said mobilizing agent is IL-3.

[0102] 98. The method of aspect 72, wherein said mobilizing agent is thrombopoietin.

[0103] 99. The method of aspect 72, wherein said mobilizing agent is hepatocyte growth factor.

[0104] 100. The method of aspect 72, wherein said mobilizing agent is FGF-1.

[0105] 101. The method of aspect 72, wherein said mobilizing agent is FGF-2.

[0106] 102. The method of aspect 72, wherein said mobilizing agent is cyclophosphamide.

[0107] 103. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from hair follicle.

[0108] 104. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from dental pulp.

[0109] 105. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from endometrial tissue..

[0110] 106. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from fallopian tubes.

[0111] 107. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from menstrual blood.

[0112] 108. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from cord blood.

[0113] 109. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from amnionic tissue.

[0114] 110. The method of aspects 56-60, wherein said mesenchymal stem cell is isolated from amnionic fluid.

[0115] 111. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interferon gamma.

[0116] 112. The method of aspect 111, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0117] 113. The method of aspect 111, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0118] 114. The method of aspect 111, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0119] 115. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TNF-alpha.

[0120] 116. The method of aspect 115, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0121] 117. The method of aspect 115, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0122] 118. The method of aspect 115, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0123] 119. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TRAIL.

[0124] 120. The method of aspect 119, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0125] 121. The method of aspect 119, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0126] 122. The method of aspect 119, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0127] 123. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to lymphotoxin.

[0128] 124. The method of aspect 123, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0129] 125. The method of aspect 123, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0130] 126. The method of aspect 123, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0131] 127. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TRANCE.

[0132] 128. The method of aspect 127, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0133] 129. The method of aspect 127, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0134] 130. The method of aspect 127, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0135] 131. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0136] 132. The method of aspect 131, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0137] 133. The method of aspect 131, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0138] 134. The method of aspect 131, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0139] 135. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0140] 136. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0141] 137. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0142] 138. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0143] 135. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0144] 136. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0145] 137. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0146] 138. The method of aspect 135, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0147] 139. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-6.

[0148] 140. The method of aspect 139, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0149] 141. The method of aspect 139, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0150] 142. The method of aspect 139, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0151] 143. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-8.

[0152] 144. The method of aspect 143, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0153] 145. The method of aspect 143, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0154] 146. The method of aspect 143, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0155] 147. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-17.

[0156] 148. The method of aspect 147, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0157] 149. The method of aspect 147, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0158] 150. The method of aspect 147, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0159] 151. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-18.

[0160] 152. The method of aspect 147, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0161] 153. The method of aspect 147, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0162] 154. The method of aspect 147, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0163] 155. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to HMGBl.

[0164] 156. The method of aspect 155, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 25% as compared to baseline.

[0165] 157. The method of aspect 155, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 50% as compared to baseline.

[0166] 158. The method of aspect 155, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase expression of interleukin-10 receptor by 100% as compared to baseline.

[0167] 159. The method of aspect 111, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of indolamine 2,3 dioxygenase by 25% as compared to baseline.

[0168] 160. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interferon gamma.

[0169] 161. The method of aspect 160, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0170] 162. The method of aspect 160, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0171] 163. The method of aspect 160, wherein said interferon gamma is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0172] 164. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TNF-alpha.

[0173] 165. The method of aspect 164, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0174] 166. The method of aspect 164, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0175] 167. The method of aspect 164, wherein said TNF-alpha is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0176] 168. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TRAIL.

[0177] 169. The method of aspect 168, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0178] 170. The method of aspect 168, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0179] 171. The method of aspect 168, wherein said TRAIL is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0180] 172. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to lymphotoxin.

[0181] 173. The method of aspect 172, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0182] 174. The method of aspect 172, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0183] 175. The method of aspect 172, wherein said lymphotoxin is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0184] 176. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to TRANCE.

[0185] 177. The method of aspect 176, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0186] 178. The method of aspect 176, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0187] 179. The method of aspect 176, wherein said TRANCE is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0188] 180. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0189] 181. The method of aspect 180, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0190] 182. The method of aspect 180, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0191] 183. The method of aspect 181, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0192] 184. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0193] 185. The method of aspect 184, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0194] 186. The method of aspect 185, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0195] 187. The method of aspect 185, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0196] 188. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-1.

[0197] 189. The method of aspect 188, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0198] 190. The method of aspect 188, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0199] 191. The method of aspect 188, wherein said interleukin-1 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0200] 192. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-6.

[0201] 193. The method of aspect 192, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0202] 194. The method of aspect 192, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0203] 195. The method of aspect 192, wherein said interleukin-6 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0204] 196. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-8.

[0205] 197. The method of aspect 196, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0206] 197. The method of aspect 196, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0207] 198. The method of aspect 196, wherein said interleukin-8 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0208] 199. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-17.

[0209] 200. The method of aspect 199, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0210] 202. The method of aspect 199, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0211] 203. The method of aspect 199, wherein said interleukin-17 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0212] 204. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to interleukin-18.

[0213] 205. The method of aspect 204, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0214] 206. The method of aspect 204, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0215] 207. The method of aspect 204, wherein said interleukin-18 is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0216] 208. The method of aspects 56-60, wherein said mesenchymal stem cell is exposed to HMGBl.

[0217] 209. The method of aspect 208, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 25% as compared to baseline.

[0218] 210. The method of aspect 208, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 50% as compared to baseline.

[0219] 211. The method of aspect 208, wherein said HMGBl is applied to said mesenchymal stem cell at a concentration and duration sufficient to increase production of LIF by 100% as compared to baseline.

[0220] 212. The method of aspect 1, wherein said representative tissue cells are tissue cells belonging to the type of tissue whose progenitors is desired to be generated.

[0221] 213. The method of aspect 212, wherein said tissue is selected from a group of tissues comprising: a) nephrotic tissue; b) hepatic tissue; c) stomach tissue; d) dermal tissue; e) thymic tissue; f) neural tissue; g) hair tissue; h) ocular tissue; i) lymphatic tissue; j) musculoskeletal tissue; k) bone tissue; I) endocrine tissue; m) vascular tissue; n) hematopoietic tissue; o) colonic tissue; p) cardiac tissue; q) muscle tissue; r) nail tissue; s) cartilage tissue; t) pancreatic tissue; u) bladder tissue; v) penile tissue; w) testicular tissue; x) prostatic tissue; y) ovarian tissue; z) fallopian tissue; aa) digestive tissue; and ab) glandular tissue.

[0222] 214. The method of aspect 213, wherein said representative tissue is created by obtaining matrix from said tissue by decellularizing said tissue and then seeding said tissue with cells.

[0223] 215. The method of aspect 214, wherein said representative tissue is treated with a cellular stressor.

[0224] 216. The method of aspect 215, wherein conditioned media is obtained from said representative tissue treated with said cellular stressor.

[0225] 217. The method of aspect 216, wherein said conditioned media is added to said pluripotent stem cell.

[0226] 218. The method of aspect 217, wherein said conditioned media is added to said pluripotent stem cell along with a "differentiation adjuvant.

[0227] 219. The method of aspect 218, wherein said differentiation adjuvant is tissue specific.

[0228] 220. The method of aspect 218, wherein said differentiation adjuvant is tissue nonspecific.

[0229] 221. The method of aspect 214, wherein said tissue is decellularized by treatment a solvent.

[0230] 222. The method of aspect 214, wherein said decellularized tissue preserves anatomical features.

[0231] 223. The method of aspect 214, wherein said decellularized tissue is seeded with pluripotent stem cells.

[0232] 224. The method of aspect 214, wherein said decellularized tissue is seeded with differentiated progenitor cells.

[0233] 225. The method of aspect 214, wherein said decellularized tissue is seeded with mesenchymal stem cells.

[0234] 226. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells.

[0235] 227. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by c-kit selection of cells from the desired tissue.

[0236] 228. The method of aspect 227, wherein said c-kit selection is performed by immunoisolation.

[0237] 229. The method of aspect 227, wherein said c-kit selection is performed by immunosurgery.

[0238] 230. The method of aspect 227 , wherein said c-kit selection is performed by magnetic activated cell sorting.

[0239] 231. The method of aspect 227 , wherein said c-kit selection is performed by fluorescent activated cell sorting.

[0240] 232. The method of aspect 227 , wherein said c-kit selection is performed by positive selection.

[0241] 233. The method of aspect 227 , wherein said c-kit selection is performed by negative selection.

[0242] 234. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by VEGF-receptor selection of cells from the desired tissue.

[0243] 235. The method of aspect 234, wherein said VEGF-receptor selection is performed by immunoisolation.

[0244] 236. The method of aspect 234, wherein said VEGF-receptor selection is performed by immunosurgery.

[0245] 237. The method of aspect 227 , wherein said VEGF-receptor selection is performed by magnetic activated cell sorting.

[0246] 238. The method of aspect 234, wherein said VEGF-receptor selection is performed by fluorescent activated cell sorting.

[0247] 239. The method of aspect 234, wherein said VEGF-receptor selection is performed by positive selection.

[0248] 240. The method of aspect 234, wherein said VEGF-receptor selection is performed by negative selection.

[0249] 241. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by aldehyde dehydrogenase selection of cells from the desired tissue.

[0250] 242. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by immunoisolation.

[0251] 243. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by immunosurgery.

[0252] 244. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by magnetic activated cell sorting.

[0253] 245. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by fluorescent activated cell sorting.

[0254] 246. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by positive selection.

[0255] 247. The method of aspect 241, wherein said aldehyde dehydrogenase selection is performed by negative selection.

[0256] 248. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by CD133 selection of cells from the desired tissue.

[0257] 249. The method of aspect 248, wherein said CD133 selection is performed by immunoisolation.

[0258] 250. The method of aspect 248, wherein said CD133 selection is performed by immunosurgery.

[0259] 251. The method of aspect 248, wherein said CD133 selection is performed by magnetic activated cell sorting.

[0260] 252. The method of aspect 248, wherein said CD133 selection is performed by fluorescent activated cell sorting.

[0261] 253. The method of aspect 248, wherein said CD133 selection is performed by positive selection.

[0262] 254. The method of aspect 248, wherein said CD133 selection is performed by negative selection.

[0263] 255. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by CD34 selection of cells from the desired tissue.

[0264] 256. The method of aspect 255, wherein said CD34 selection is performed by immunoisolation.

[0265] 257. The method of aspect 255, wherein said CD34 selection is performed by immunosurgery.

[0266] 258. The method of aspect 255, wherein said CD34 selection is performed by magnetic activated cell sorting.

[0267] 259. The method of aspect 255, wherein said CD34 selection is performed by fluorescent activated cell sorting.

[0268] 260. The method of aspect 255, wherein said CD34 selection is performed by positive selection.

[0269] 261. The method of aspect 255, wherein said CD34 selection is performed by negative selection.

[0270] 262. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by CD73 selection of cells from the desired tissue.

[0271] 263. The method of aspect 262, wherein said CD73 selection is performed by immunoisolation.

[0272] 264. The method of aspect 262, wherein said CD73 selection is performed by immunosurgery.

[0273] 265. The method of aspect 262, wherein said CD73 selection is performed by magnetic activated cell sorting.

[0274] 266. The method of aspect 262, wherein said CD73 selection is performed by fluorescent activated cell sorting.

[0275] 267. The method of aspect 262, wherein said CD73 selection is performed by positive selection.

[0276] 268. The method of aspect 262, wherein said CD73 selection is performed by negative selection.

[0277] 269. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by c-met selection of cells from the desired tissue.

[0278] 270. The method of aspect 269, wherein said c-met selection is performed by immunoisolation.

[0279] 271. The method of aspect 269, wherein said c-met selection is performed by immunosurgery.

[0280] Til. The method of aspect 269, wherein said c-met selection is performed by magnetic activated cell sorting.

[0281] 273. The method of aspect 269, wherein said c-met selection is performed by fluorescent activated cell sorting.

[0282] 274. The method of aspect 269, wherein said c-met selection is performed by positive selection.

[0283] 275. The method of aspect 269, wherein said c-met selection is performed by negative selection.

[0284] 276. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by PD-L1 selection of cells from the desired tissue.

[0285] Til . The method of aspect 276, wherein said PD-L1 selection is performed by immunoisolation.

[0286] 278. The method of aspect 276, wherein said PD-L1 selection is performed by immunosurgery.

[0287] 279. The method of aspect 276, wherein said PD-L1 selection is performed by magnetic activated cell sorting.

[0288] 280. The method of aspect 276, wherein said PD-L1 selection is performed by fluorescent activated cell sorting.

[0289] 281. The method of aspect 276, wherein said PD-L1 selection is performed by positive selection.

[0290] 282. The method of aspect 276, wherein said PD-L1 selection is performed by negative selection.

[0291] 283. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by c-mpl selection of cells from the desired tissue.

[0292] 284. The method of aspect 283, wherein said c-mpl selection is performed by immunoisolation.

[0293] 285. The method of aspect 283, wherein said c-mpl selection is performed by immunosurgery.

[0294] 286. The method of aspect 283, wherein said c-mpl selection is performed by magnetic activated cell sorting.

[0295] 287. The method of aspect 283, wherein said c-mpl selection is performed by fluorescent activated cell sorting.

[0296] 288. The method of aspect 283, wherein said c-mpl selection is performed by positive selection.

[0297] 289. The method of aspect 283, wherein said c-mpl selection is performed by negative selection.

[0298] 290. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by pim-1 selection of cells from the desired tissue.

[0299] 291. The method of aspect 290, wherein said pim-1 selection is performed by immunoisolation.

[0300] 292. The method of aspect 290, wherein said pim-1 selection is performed by immunosurgery.

[0301] 293. The method of aspect 290, wherein said pim-1 selection is performed by magnetic activated cell sorting.

[0302] 294. The method of aspect 290, wherein said pim-1 selection is performed by fluorescent activated cell sorting.

[0303] 295. The method of aspect 290, wherein said pim-1 selection is performed by positive selection.

[0304] 296. The method of aspect 290, wherein said pim-1 selection is performed by negative selection.

[0305] 297. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by interleukin 1 receptor selection of cells from the desired tissue.

[0306] 298. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by immunoisolation.

[0307] 299. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by immunosurgery.

[0308] 300. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by magnetic activated cell sorting.

[0309] 301. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by fluorescent activated cell sorting.

[0310] 302. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by positive selection.

[0311] 303. The method of aspect 297, wherein said interleukin 1 receptor selection is performed by negative selection.

[0312] 304. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by interleukin 6 receptor selection of cells from the desired tissue.

[0313] 305. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by immunoisolation.

[0314] 306. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by immunosurgery.

[0315] 307. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by magnetic activated cell sorting.

[0316] 308. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by fluorescent activated cell sorting.

[0317] 309. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by positive selection.

[0318] 310. The method of aspect 304, wherein said interleukin 6 receptor selection is performed by negative selection.

[0319] 311. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by interleukin 3 receptor selection of cells from the desired tissue.

[0320] 312. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by immunoisolation.

[0321] 313. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by immunosurgery.

[0322] 314. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by magnetic activated cell sorting.

[0323] 315. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by fluorescent activated cell sorting.

[0324] 316. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by positive selection.

[0325] 317. The method of aspect 311, wherein said interleukin 3 receptor selection is performed by negative selection.

[0326] 318. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by interleukin 11 receptor selection of cells from the desired tissue.

[0327] 319. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by immunoisolation.

[0328] 320. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by immunosurgery.

[0329] 321. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by magnetic activated cell sorting.

[0330] 322. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by fluorescent activated cell sorting.

[0331] 323. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by positive selection.

[0332] 324. The method of aspect 318, wherein said interleukin 11 receptor selection is performed by negative selection.

[0333] 325. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by EGF receptor selection of cells from the desired tissue.

[0334] 326. The method of aspect 325, wherein said EGF receptor selection is performed by immunoisolation.

[0335] 327. The method of aspect 325, wherein said EGF receptor selection is performed by immunosurgery.

[0336] 328. The method of aspect 325, wherein said EGF receptor selection is performed by magnetic activated cell sorting.

[0337] 329. The method of aspect 325, wherein said EGF receptor selection is performed by fluorescent activated cell sorting.

[0338] 330. The method of aspect 325, wherein said EGF receptor selection is performed by positive selection.

[0339] 331. The method of aspect 325, wherein said EGF receptor selection is performed by negative selection.

[0340] 332. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by PDGF receptor selection of cells from the desired tissue.

[0341] 333. The method of aspect 332, wherein said PDGF receptor selection is performed by immunoisolation.

[0342] 334. The method of aspect 332, wherein said PDGF receptor selection is performed by immunosurgery.

[0343] 335. The method of aspect 332, wherein said PDGF receptor selection is performed by magnetic activated cell sorting.

[0344] 336. The method of aspect 332, wherein said PDGF receptor selection is performed by fluorescent activated cell sorting.

[0345] 337. The method of aspect 332, wherein said PDGF receptor selection is performed by positive selection.

[0346] 338. The method of aspect 332, wherein said PDGF receptor selection is performed by negative selection.

[0347] 339. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by FGF-1 receptor selection of cells from the desired tissue.

[0348] 340. The method of aspect 339, wherein said FGF-1 receptor selection is performed by immunoisolation.

[0349] 341. The method of aspect 339, wherein said FGF-1 receptor selection is performed by immunosurgery.

[0350] 342. The method of aspect 339, wherein said FGF-1 receptor selection is performed by magnetic activated cell sorting.

[0351] 343. The method of aspect 339, wherein said FGF-1 receptor selection is performed by fluorescent activated cell sorting.

[0352] 344. The method of aspect 339, wherein said FGF-1 receptor selection is performed by positive selection.

[0353] 345. The method of aspect 339, wherein said FGF-1 receptor selection is performed by negative selection.

[0354] 346. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by FGF-1 receptor selection of cells from the desired tissue.

[0355] 347. The method of aspect 346, wherein said FGF-1 receptor selection is performed by immunoisolation.

[0356] 348. The method of aspect 346, wherein said FGF-1 receptor selection is performed by immunosurgery.

[0357] 349. The method of aspect 346, wherein said FGF-1 receptor selection is performed by magnetic activated cell sorting.

[0358] 350. The method of aspect 346, wherein said FGF-1 receptor selection is performed by fluorescent activated cell sorting.

[0359] 351. The method of aspect 346, wherein said FGF-1 receptor selection is performed by positive selection.

[0360] 352. The method of aspect 346, wherein said FGF-1 receptor selection is performed by negative selection.

[0361] 353. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by FGF-2 receptor selection of cells from the desired tissue.

[0362] 354. The method of aspect 353, wherein said FGF-2 receptor selection is performed by immunoisolation.

[0363] 355. The method of aspect 353, wherein said FGF-2 receptor selection is performed by immunosurgery.

[0364] 356. The method of aspect 353, wherein said FGF-2 receptor selection is performed by magnetic activated cell sorting.

[0365] 357. The method of aspect 353, wherein said FGF-2 receptor selection is performed by fluorescent activated cell sorting.

[0366] 358. The method of aspect 353, wherein said FGF-2 receptor selection is performed by positive selection.

[0367] 359. The method of aspect 353, wherein said FGF-2 receptor selection is performed by negative selection.

[0368] 360. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by galectin-3 receptor selection of cells from the desired tissue.

[0369] 361. The method of aspect 360, wherein said galectin-3 receptor selection is performed by immunoisolation.

[0370] 362. The method of aspect 361, wherein said galectin-3 receptor selection is performed by immunosurgery.

[0371] 363. The method of aspect 361, wherein said galectin-3 receptor selection is performed by magnetic activated cell sorting.

[0372] 364. The method of aspect 361, wherein said galectin-3 receptor selection is performed by fluorescent activated cell sorting.

[0373] 365. The method of aspect 361, wherein said galectin-3 receptor selection is performed by positive selection.

[0374] 366. The method of aspect 361, wherein said galectin-3 receptor selection is performed by negative selection.

[0375] 367. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by Jagged receptor selection of cells from the desired tissue.

[0376] 368. The method of aspect 367, wherein said Jagged receptor selection is performed by immunoisolation.

[0377] 369. The method of aspect 367, wherein said Jagged receptor selection is performed by immunosurgery.

[0378] 370. The method of aspect 367, wherein said Jagged receptor selection is performed by magnetic activated cell sorting.

[0379] 371. The method of aspect 367, wherein said Jagged receptor selection is performed by fluorescent activated cell sorting.

[0380] 372. The method of aspect 367, wherein said Jagged receptor selection is performed by positive selection.

[0381] 373. The method of aspect 367, wherein said Jagged receptor selection is performed by negative selection.

[0382]

[0383] 374. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by GDF-11 receptor selection of cells from the desired tissue.

[0384] 375. The method of aspect 374, wherein said GDF-11 receptor selection is performed by immunoisolation.

[0385] 376. The method of aspect 374, wherein said GDF-11 receptor selection is performed by immunosurgery.

[0386] 377. The method of aspect 374, wherein said GDF-11 receptor selection is performed by magnetic activated cell sorting.

[0387] 378. The method of aspect 374, wherein said GDF-11 receptor selection is performed by fluorescent activated cell sorting.

[0388] 379. The method of aspect 374, wherein said GDF-11 receptor selection is performed by positive selection.

[0389] 380. The method of aspect 374, wherein said GDF-11 receptor selection is performed by negative selection.

[0390] 381. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by angiopoietin receptor selection of cells from the desired tissue. 1

[0391] 382. The method of aspect 381, wherein said angiopoietin receptor selection is performed by immunoisolation.

[0392] 383. The method of aspect 381, wherein said angiopoietin receptor selection is performed by immunosurgery.

[0393] 384. The method of aspect 381, wherein said angiopoietin receptor selection is performed by magnetic activated cell sorting.

[0394] 385. The method of aspect 381, wherein said angiopoietin receptor selection is performed by fluorescent activated cell sorting.

[0395] 386. The method of aspect 381, wherein said angiopoietin receptor selection is performed by positive selection.

[0396] 387. The method of aspect 381, wherein said angiopoietin receptor selection is performed by negative selection.

[0397] 388. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by TGF-beta receptor selection of cells from the desired tissue.

[0398] 389. The method of aspect 388, wherein said TGF-beta receptor selection is performed by immunoisolation.

[0399] 390. The method of aspect 388, wherein said TGF-beta receptor selection is performed by immunosurgery.

[0400] 391. The method of aspect 388, wherein said TGF-beta receptor selection is performed by magnetic activated cell sorting.

[0401] 392. The method of aspect 388, wherein said TGF-beta receptor selection is performed by fluorescent activated cell sorting.

[0402] 393. The method of aspect 388, wherein said TGF-beta receptor selection is performed by positive selection.

[0403] 394. The method of aspect 388, wherein said TGF-beta receptor selection is performed by negative selection.

[0404] 395. The method of aspect 214, wherein said decellularized tissue is seeded with tissue specific progenitor cells isolated by endoglin receptor selection of cells from the desired tissue.

[0405] 396. The method of aspect 395, wherein said endoglin receptor selection is performed by immunoisolation.

[0406] 397. The method of aspect 395, wherein said endoglin receptor selection is performed by immunosurgery.

[0407] 398. The method of aspect 395, wherein said endoglin receptor selection is performed by magnetic activated cell sorting.

[0408] 399. The method of aspect 395, wherein said endoglin receptor selection is performed by fluorescent activated cell sorting.

[0409] 400. The method of aspect 395, wherein said endoglin receptor selection is performed by positive selection.

[0410] 401. The method of aspect 395, wherein said endoglin receptor selection is performed by negative selection.

[0411] 402. The method of aspects 227 to 401, wherein said tissue specific progenitors are expanded without substantial differentiation.

[0412] 403. The method of aspect 402, wherein said expansion of tissue specific progenitors is performed without substantial differentiation by culture in the presence of one or moreg ROCK inhibitor.

[0413] 404. The method of aspect 402, wherein said expansion of tissue specific progenitors is performed without substantial differentiation by culture in the presence of one or more histone deacetylase inhibitor.

[0414] 405. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded in the presence of means capable of enhancing viability of said cells.

[0415] 406. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hypoxia.

[0416] 407. The method of aspect 406, wherein said cells are exposed to an atmosphere of 0.1-4% oxygen for a period of time of 5 minutes to 8 hours.

[0417] 408. The method of aspect 406, wherein said cells are exposed to an atmosphere of 0.5-4% oxygen for a period of time of 5 minutes to 8 hours.

[0418] 409. The method of aspect 406, wherein said cells are exposed to an atmosphere of 1-4% oxygen for a period of time of 5 minutes to 8 hours.

[0419] 410. The method of aspect 406, wherein said cells are exposed to an atmosphere of 2-4% oxygen for a period of time of 5 minutes to 8 hours.

[0420] 411. The method of aspect 406, wherein said cells are exposed to an atmosphere of 2-4% oxygen for a period of time of 1 to 8 hours.

[0421] 412. The method of aspect 406, wherein said cells are exposed to an atmosphere of 2-4% oxygen for a period of time of 2 to 8 hours.

[0422] 413. The method of aspect 406, wherein said cells are exposed to an atmosphere of 2-4% oxygen for a period of time of 4 to 8 hours.

[0423] 414. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2 by 10% or more as compared to baseline.

[0424] 415. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2 by 20% or more as compared to baseline.

[0425] 416. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2 by 100% or more as compared to baseline.

[0426] 417. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of survivin by 10% or more as compared to baseline.

[0427] 418. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of survivin by 20% or more as compared to baseline.

[0428] 419. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of survivin by 100% or more as compared to baseline.

[0429] 420. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of livin by 10% or more as compared to baseline.

[0430] 421. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of livin by 20% or more as compared to baseline.

[0431] 339. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of livin by 100% or more as compared to baseline.

[0432] 340. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of IAP-1 by 10% or more as compared to baseline.

[0433] 341. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of lAP-lby 20% or more as compared to baseline.

[0434] 342. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of lAP-lby 100% or more as compared to baseline.

[0435] 343. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2xL by 10% or more as compared to baseline.

[0436] 344. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2xL by 20% or more as compared to baseline.

[0437] 345. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hydrogen gas at a concentration and time period sufficient to increase levels of bcl-2xL by 100% or more as compared to baseline.

[0438] 346. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2 by 10% or more as compared to baseline.

[0439] 347. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2 by 20% or more as compared to baseline.

[0440] 348. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2 by 100% or more as compared to baseline.

[0441] 349. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of survivin by 10% or more as compared to baseline.

[0442] 350. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of survivin by 20% or more as compared to baseline.

[0443] 351. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of survivin by 100% or more as compared to baseline.

[0444] 352. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of livin by 10% or more as compared to baseline.

[0445] 353. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of livin by 20% or more as compared to baseline.

[0446] 354. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of livin by 100% or more as compared to baseline.

[0447] 355. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of IAP-1 by 10% or more as compared to baseline.

[0448] 356. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of lAP-lby 20% or more as compared to baseline.

[0449] 357. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of lAP-lby 100% or more as compared to baseline.

[0450] 358. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2xL by 10% or more as compared to baseline.

[0451] 359. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2xL by 20% or more as compared to baseline.

[0452] 360. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with argon gas at a concentration and time period sufficient to increase levels of bcl-2xL by 100% or more as compared to baseline.

[0453] 361. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2 by 10% or more as compared to baseline.

[0454] 362. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2 by 20% or more as compared to baseline.

[0455] 363. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2 by 100% or more as compared to baseline.

[0456] 364. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of survivin by 10% or more as compared to baseline.

[0457] 365. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of survivin by 20% or more as compared to baseline.

[0458] 366. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of survivin by 100% or more as compared to baseline.

[0459] 367. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of livin by 10% or more as compared to baseline.

[0460] 368. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of livin by 20% or more as compared to baseline.

[0461] 369. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of livin by 100% or more as compared to baseline.

[0462] 370. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of IAP-1 by 10% or more as compared to baseline.

[0463] 371. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of lAP-lby 20% or more as compared to baseline.

[0464] 372. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of lAP-lby 100% or more as compared to baseline.

[0465] 373. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2xL by 10% or more as compared to baseline.

[0466] 374. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2xL by 20% or more as compared to baseline.

[0467] 375. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with n-acetylcysteine at a concentration and time period sufficient to increase levels of bcl-2xL by 100% or more as compared to baseline.

[0468] 376. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2 by 10% or more as compared to baseline.

[0469] 377. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2 by 20% or more as compared to baseline.

[0470] 378. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2 by 100% or more as compared to baseline.

[0471] 379. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of survivin by 10% or more as compared to baseline.

[0472] 380. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of survivin by 20% or more as compared to baseline.

[0473] 381. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of survivin by 100% or more as compared to baseline.

[0474] 382. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of livin by 10% or more as compared to baseline.

[0475] 383. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of livin by 20% or more as compared to baseline.

[0476] 384. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of livin by 100% or more as compared to baseline.

[0477] 385. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of IAP-1 by 10% or more as compared to baseline.

[0478] 386. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of lAP-lby 20% or more as compared to baseline.

[0479] 387. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of lAP-lby 100% or more as compared to baseline.

[0480] 388. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2xL by 10% or more as compared to baseline.

[0481] 389. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2xL by 20% or more as compared to baseline.

[0482] 390. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with erythropoietin at a concentration and time period sufficient to increase levels of bcl-2xL by 100% or more as compared to baseline.

[0483] 391. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with FGF-1.

[0484] 392. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Z-VAD-FMK.

[0485] 393. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Z-VAD- (OME)-FMK.

[0486] 394. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Q-VD-OPH.

[0487] 395. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Belnacasan.

[0488] 396. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Z-DEVD-FMK.

[0489] 397. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with Ac-DEVD-FMK.

[0490] 398. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with dehydrocorydaline.

[0491] 399. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with 714-X.

[0492] 400. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting bcl-2Xs.

[0493] 401. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting bcl-2Xs.

[0494] 402. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting bcl-2Xs.

[0495] 403. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of bcl-2Xs.

[0496] 404. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting caspase-3.

[0497] 405. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting caspase-3.

[0498] 406. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting caspase-3.

[0499] 407. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of caspase-3.

[0500] 408. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting caspase-.8

[0501] 409. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting caspase-8.

[0502] 410. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting caspase-8.

[0503] 411. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of caspase-8.

[0504] 412. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting caspase-9

[0505] 413. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting caspase-9.

[0506] 414. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting caspase-9.

[0507] 415. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of caspase-9.

[0508] 416. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting Bax.

[0509] 417. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting Bax.

[0510] 418. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting Bax.

[0511] 419. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of Bax.

[0512] 420. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting Bid.

[0513] 421. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting Bid.

[0514] 422. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting Bid.

[0515] 423. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of Bid.

[0516] 424. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with siRNA targeting Hrk.

[0517] 425. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with antisense oligonucleotide targeting Hrk.

[0518] 426. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with hammerhead ribozyme targeting Hrk.

[0519] 427. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with gene edit targeting of Hrk.

[0520] 428. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with a stimulator of NF-kappa B.

[0521] 429. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with minocycline.

[0522] 430. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with doxycycline.

[0523] 431. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with creatinine.

[0524] 432. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with dicholoroacetate.

[0525] 433. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with coenzyme Q10.

[0526] 434. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with lipoic acid.

[0527] 435. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with retinoic acid.

[0528] 436. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with all trans retinoic acid.

[0529] 437. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with vitamin D3.

[0530] 438. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with cystamine.

[0531] 439. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with bucellamine.

[0532] 440. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with riluozole.

[0533] 441. The method of aspect 405, wherein said enhancement of viability is elicited by pretreatment of cells with linderalactone.

[0534] 442. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by administration of a histone deacetylase inhibitor.

[0535] 443. The method of aspect 442, wherein said histone deacetylase inhibitor is siRNA targeting one or more histone deacetylases.

[0536] 444. The method of aspect 442, wherein said histone deacetylase inhibitor is microRNA targeting one or more histone deacetylases.

[0537] 445. The method of aspect 442, wherein said histone deacetylase inhibitor is antisense oligonucleotides targeting one or more histone deacetylases.

[0538] 446. The method of aspect 442, wherein said histone deacetylase inhibitor is decoy oligonucleotides targeting one or more histone deacetylases.

[0539] 447. The method of aspect 442, wherein said histone deacetylase inhibitor is hammerhead ribozymes targeting one or more histone deacetylases.

[0540] 448. The method of aspect 442, wherein said histone deacetylase inhibitor is sulforaphane.

[0541] 449. The method of aspect 442, wherein said histone deacetylase inhibitor is valproic acid.

[0542] 450. The method of aspect 442, wherein said histone deacetylase inhibitor is trichostatin A.

[0543] 451. The method of aspect 442, wherein said histone deacetylase inhibitor is phenylbutyrate.

[0544] 452. The method of aspect 442, wherein said histone deacetylase inhibitor is vorinostat.

[0545] 453. The method of aspect 442, wherein said histone deacetylase inhibitor isRomidepsin.

[0546] 454. The method of aspect 442, wherein said histone deacetylase inhibitor isPanobinostat.

[0547] 455. The method of aspect 442, wherein said histone deacetylase inhibitor isBelinostat

[0548] 456. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with an inhibitor of NF-kappa B.

[0549] 457. The method of aspect 456, wherein said NF-kappa B inhibitor is n- acetylcysteine.

[0550]

[0551] 458. The method of aspect 456, wherein said NF-kappa B inhibitor is ascorbic acid.

[0552] 459. The method of aspect 456, wherein said NF-kappa B inhibitor is alpha lipoic acid.

[0553] 460. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with a growth factor.

[0554] 461. The method of aspect 460, wherein said growth factor is FGF-1.

[0555] 462. The method of aspect 460, wherein said growth factor FGF-2.

[0556] 463. The method of aspect 460, wherein said growth factor is FGF-5.

[0557] 464. The method of aspect 460, wherein said growth factor PDGF.

[0558] 465. The method of aspect 460, wherein said growth factor is PDGF-BB.

[0559] 466. The method of aspect 460, wherein said growth factor angiopoietin.

[0560] 467. The method of aspect 460, wherein said growth factor is cobra venom factor.

[0561] 468. The method of aspect 460, wherein said growth factor is complement component Clq.

[0562] 469. The method of aspect 460, wherein said growth factor is complement component C3a.

[0563] 470. The method of aspect 460, wherein said growth factor is complement component C5a.

[0564] 471. The method of aspect 460, wherein said growth factor is VEGF.

[0565] 472. The method of aspect 460, wherein said growth factor is VEGF-C.

[0566] 473. The method of aspect 460, wherein said growth factor is EGF.

[0567] 474. The method of aspect 460, wherein said growth factor is FGF-1.

[0568] 475. The method of aspect 460, wherein said growth factor is FGF-2.

[0569] 476. The method of aspect 460, wherein said growth factor is FGF-5.

[0570] 477. The method of aspect 460, wherein said growth factor is IGF.

[0571] 478. The method of aspect 460, wherein said growth factor is Klotho.

[0572] 479. The method of aspect 460, wherein said growth factor is angiopoietin.

[0573] 480. The method of aspect 460, wherein said growth factor is TGF-beta.

[0574] 481. The method of aspect 460, wherein said growth factor is endoglin.

[0575] 482. The method of aspect 460, wherein said growth factor is NGF.

[0576] 483. The method of aspect 460, wherein said growth factor is HGF.

[0577] 484. The method of aspect 460, wherein said growth factor is insulin.

[0578] 485. The method of aspect 460, wherein said growth factor is erythropoietin.

[0579] 486. The method of aspect 460, wherein said growth factor is protein C.

[0580] 487. The method of aspect 460, wherein said growth factor is activated proteinC.

[0581] 488. The method of aspect 460, wherein said growth factor is insulin growth factor binding protein.

[0582] 489. The method of aspect 460, wherein said growth factor is somatostatin.

[0583] 490. The method of aspect 460, wherein said growth factor is annexin-V.

[0584] 491. The method of aspect 460, wherein said growth factor is IL-3.

[0585] 492. The method of aspect 460, wherein said growth factor is IL-4.

[0586] 493. The method of aspect 460, wherein said growth factor is IL-6.

[0587] 494. The method of aspect 460, wherein said growth factor is IL-8.

[0588] 495. The method of aspect 460, wherein said growth factor is IL-10.

[0589] 496. The method of aspect 460, wherein said growth factor is IL-12.

[0590] 497. The method of aspect 460, wherein said growth factor is IL-20.

[0591] 498. The method of aspect 460, wherein said growth factor is IL-22.

[0592] 499. The method of aspect 460, wherein said growth factor is IL-37.

[0593] 500. The method of aspect 460, wherein said growth factor is IL-38.

[0594] 501. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with MMP3.

[0595] 502. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with MMP5.

[0596] 503. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with MMP7.

[0597] 504. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with MMP9.

[0598] 505. The method of aspects 225 to 401, wherein said tissue specific progenitors are expanded by treatment with MMP13.

[0599] 506. A method of generating cardiac specific progenitor cells through the steps of: a) obtaining a pluripotent stem cell population; b) treating said pluripotent stem cell population with one or more cardiogenic media; c) extracting cardiac progenitor cells obtained from culture of said pluripotent stem cell population treated with said cardiogenic media; d) expanding said cardiac progenitor cells; and e) cryogenically banking said cardiac progenitor cells.

[0600] 507. The method of aspect 1, wherein said cardiogenic media is conditioned media from a cardiac progenitor cell line.

[0601] 508. The method of aspect 507, wherein said cardiac progenitor cell line is immortalized.

[0602] 509. The method of aspect 508, wherein said immortalization is accomplished by treatment agents that increase expression of hTERT.

[0603] 510. The method of aspect 509, wherein said treatment that increases expression of hTERT is followed by transfection with one or more immortalization genes.

[0604] 511. The method of aspect 510, wherein said hTERT increasing treatment is hypoxia.

[0605] 512. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF-1 by more than 25% compared to baseline.

[0606] 513. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF-1 by more than 50% compared to baseline.

[0607] 514. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF-1 by more than 100% compared to baseline.

[0608] 515. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-1 by more than 25% compared to baseline.

[0609] 516. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-1 by more than 50% compared to baseline.

[0610] 517. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-1 by more than 100% compared to baseline.

[0611] 518. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-2 by more than 25% compared to baseline.

[0612] 519. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-2 by more than 50% compared to baseline.

[0613] 520. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of FGF-2 by more than 100% compared to baseline.

[0614] 521. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of EGF by more than 25% compared to baseline.

[0615] 522. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of EGF by more than 50% compared to baseline.

[0616] 523. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of EGF by more than 100% compared to baseline.

[0617] 524. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of amphiregulin by more than 25% compared to baseline.

[0618] 525. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of amphiregulin by more than 50% compared to baseline.

[0619] 526. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of amphiregulin by more than 100% compared to baseline.

[0620] 527. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TGF-beta by more than 25% compared to baseline.

[0621] 528. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TGF-beta by more than 50% compared to baseline.

[0622] 529. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TGF-beta by more than 100% compared to baseline.

[0623] 530. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of endoglin by more than 25% compared to baseline.

[0624] 531. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of endoglin by more than 50% compared to baseline.

[0625] 532. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of endoglin by more than 100% compared to baseline.

[0626] 533. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of VEGF by more than 25% compared to baseline.

[0627] 534. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of VEGF by more than 50% compared to baseline.

[0628] 535. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of VEGF by more than 100% compared to baseline.

[0629] 536. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IGF by more than 25% compared to baseline.

[0630] 537. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IGF by more than 50% compared to baseline.

[0631] 538. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IGF by more than 100% compared to baseline.

[0632] 539. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-1 by more than 25% compared to baseline.

[0633] 540. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-lby more than 50% compared to baseline.

[0634] 541. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-lby more than 100% compared to baseline.

[0635] 542. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-3 by more than 25% compared to baseline.

[0636] 543. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-3 by more than 50% compared to baseline.

[0637] 544. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-3 by more than 100% compared to baseline.

[0638] 545. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-9 by more than 25% compared to baseline.

[0639] 546. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-9 by more than 50% compared to baseline.

[0640] 547. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of galectin-9 by more than 100% compared to baseline.

[0641] 548. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-3 by more than 25% compared to baseline.

[0642] 549. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-3 by more than 50% compared to baseline.

[0643] 550. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-3 by more than 100% compared to baseline.

[0644] 551. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-9 by more than 25% compared to baseline.

[0645] 552. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-9 by more than 50% compared to baseline.

[0646] 553. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of MMP-9 by more than 100% compared to baseline.

[0647] 554. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of angiopoietin by more than 25% compared to baseline.

[0648] 555. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of angiopoietin by more than 50% compared to baseline.

[0649] 556. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of angiopoietin by more than 100% compared to baseline.

[0650] 557. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of PDGF-BB by more than 25% compared to baseline.

[0651] 558. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of PDGF-BB by more than 50% compared to baseline.

[0652] 559. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of PDGF-BB by more than 100% compared to baseline.

[0653] 560. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF by more than 25% compared to baseline.

[0654] 561. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF by more than 50% compared to baseline.

[0655] 562. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HGF by more than 100% compared to baseline.

[0656] 563. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of soluble HLA-G by more than 25% compared to baseline.

[0657] 564. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of soluble HLA-G by more than 50% compared to baseline.

[0658] 565. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of soluble HLA-G by more than 100% compared to baseline.

[0659] 566. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of NRF2 by more than 25% compared to baseline.

[0660] 567. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of NRF2 by more than 50% compared to baseline.

[0661] 568. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of NRF2 by more than 100% compared to baseline.

[0662] 569. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HO-1 by more than 25% compared to baseline.

[0663] 570. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HO-1 by more than 50% compared to baseline.

[0664] 571. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of HO-1 by more than 100% compared to baseline.

[0665] 572. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2 by more than 25% compared to baseline.

[0666] 573. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2 by more than 50% compared to baseline.

[0667] 574. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2 by more than 100% compared to baseline.

[0668] 575. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2xL by more than 25% compared to baseline.

[0669] 576. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2xL by more than 50% compared to baseline.

[0670] 577. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of bcl-2xL by more than 100% compared to baseline.

[0671] 578. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of livin by more than 25% compared to baseline.

[0672] 579. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of livin by more than 50% compared to baseline.

[0673] 580. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of livin by more than 100% compared to baseline.

[0674] 581. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of survivin by more than 25% compared to baseline.

[0675] 582. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of survivin by more than 50% compared to baseline.

[0676] 583. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of survivin by more than 100% compared to baseline.

[0677] 584. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of c-met by more than 25% compared to baseline.

[0678] 585. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of c-met by more than 50% compared to baseline.

[0679] 586. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of c-met by more than 100% compared to baseline.

[0680] 587. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of Al-l by more than 25% compared to baseline.

[0681] 588. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of Al-l by more than 50% compared to baseline.

[0682] 589. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of Al-l by more than 100% compared to baseline.

[0683] 590. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TAP-1 by more than 25% compared to baseline.

[0684] 591. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TAP-1 by more than 50% compared to baseline.

[0685] 592. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of TAP-1 by more than 100% compared to baseline.

[0686] 593. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IL-10 receptor by more than 25% compared to baseline.

[0687] 594. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IL-10 receptor by more than 50% compared to baseline.

[0688] 595. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of IL-10 receptor by more than 100% compared to baseline.

[0689] 596. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of arginase by more than 25% compared to baseline.

[0690] 597. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of arginase by more than 50% compared to baseline.

[0691] 598. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of arginase by more than 100% compared to baseline.

[0692] 599. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of indolamine 2,3 dioxygenase by more than 25% compared to baseline.

[0693] 600. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of indolamine 2,3 dioxygenase by more than 50% compared to baseline.

[0694] 601. The method of aspect 511, wherein said hypoxia is applied for a sufficient time and intensity to stimulate production of indolamine 2,3 dioxygenase by more than 100% compared to baseline.

[0695] 602. The method of aspect 510, wherein said immortalizing gene is the large T antigen.

[0696] 603. The method of aspect 510, wherein said immortalizing gene is PIMl.

[0697] 604. The method of aspect 510, wherein said immortalizing gene is Ras.

[0698] 605. The method of aspect 510, wherein said immortalizing gene is Raf.

[0699] 606. The method of aspect 510, wherein said immortalizing gene is abl.

[0700] 607. The method of aspect 510, wherein said immortalizing gene is bcr-abl.

[0701] 608. The method of aspect 510, wherein said immortalizing gene is vav.

[0702] 609. The method of aspect 510, wherein said immortalizing gene is erb.

[0703] 610. The method of aspect 510, wherein said immortalizing gene is her2.

[0704] 611. The method of aspect 510, wherein said immortalizing gene is c-kit.

[0705] 612. The method of aspect 510, wherein said immortalizing genes are transferred to said target cell by fusion with a neoplastically transformed cell.

[0706] 613. The method of aspect 510, wherein said immortalizing genes are activated in a temperature dependent manner.

[0707] 614. The method of aspect 56, wherein said pluripotent stem cell is generated form a partially undifferentiated stem cell.

[0708] 615. The method of aspect 614, wherein said partially undifferentiated stem cell is a mesenchymal stem cell.

[0709] 616. The metho of aspect 615, wherein said partially undifferentiated stem cell is generated by undifferentiating a monocyte or monocytic cell.

[0710] 617. The method of aspect 616, wherein said monocytic cell expresses c-kit.

[0711] 618. The method of aspect 616, wherein said monocytic cell expresses CD16.

[0712] 619. The method of aspect 616, wherein said monocytic cell expresses CD14.

[0713] 620. The method of aspect 616, wherein said monocytic cell expresses arginase.

[0714] 621. The method of aspect 616, wherein said monocytic cell expresses CD246.

[0715] 622. The method of aspect 616, wherein said monocytic cell is treated with a histone deacetylase inhibitor.

[0716] 623. The method of aspect 622, wherein said monocytic cell is transfected with a dedifferentiating agent.

[0717] 624. The method of aspect 623, wherein said dedifferentiating agent is KLF4.

[0718] 625. The method of aspect 623, wherein said dedifferentiating agent is c-myc.

[0719] 626. The method of aspect 623, wherein said dedifferentiating agent is KLF-4, c- myc, and TGF-beta receptor.

[0720] 627. The method of aspect 623, wherein said dedifferentiating agent is IL-3 receptor.

[0721] 628. The method of aspect 623, w9herein said dedifferentiating agent is IL-3 receptor and KLR-4.

[0722] 629. The method of aspect 623, wherein said dedifferentiating agent is IL-3 receptor, KLR-4 and c-myc.

[0723] 630. The method of aspect 623, wherein said dedifferentiating agent is IL-3 receptor, KLR-4, c-myc and TGF-beta receptor.

[0724] 631. The method of aspect 623, wherein said dedifferentiating agent is hTERT.

[0725] 632. The method of aspect 623, wherein said dedifferentiating agent is OCT4.

[0726] 633. The method of aspect 623, wherein said dedifferentiating agent is NANOG.

[0727] 634. The method of aspect 623, wherein said dedifferentiating agent is NANOG and OCT4.

[0728] 635. The method of aspect 623, wherein said dedifferentiating agent is cytoplasm from a pluripotent stem cell.

[0729] 636. The method of aspect 623, wherein said dedifferentiating agent is exosomes from a pluripotent stem cell.

[0730] 637. The method of aspect 623, wherein said dedifferentiating agent is miRNA from a pluripotent stem cell.

[0731] 638. The method of aspect 635 to 637, wherein said pluripotent stem cell is an iPSC cell.

[0732] 639. The method of aspect 635 to 637, wherein said pluripotent stem cell is a parthenogenesis derived stem cell.

[0733] 640. The method of aspect 635 to 637, wherein said pluripotent stem cell is a somatic cell nuclear transfer derived stem cell.

[0734] 641. The method of aspect 635 to 637, wherein said pluripotent stem cell is an iPSC cell.

[0735] 642. The method of aspect 622, wherein said histone deacetylase inhibitor is valproic acid.

[0736] 643. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0737] 644. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0738] 645. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0739] 646. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0740] 647. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0741] 648. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0742] 649. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0743] 650. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0744] 651. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0745] 652. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0746] 653. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0747] 654. The method of aspect 642, wherein said valproic acid is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0748] 655. The method of aspect 622, wherein said histone deacetylase inhibitor is trichostatin A.

[0749] 656. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0750] 657. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0751] 658. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0752] 659. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0753] 660. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0754] 661. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0755] 662. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0756] 663. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0757] 664. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0758] 665. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0759] 666. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0760] 667. The method of aspect 655, wherein said trichostatin A is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0761] 668. The method of aspect 622, wherein said histone deacetylase inhibitor is sulforaphane.

[0762] 669. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0763] 670. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0764] 671. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0765] 672. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0766] 673. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0767] 674. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0768] 675. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0769] 676. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0770] 677. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0771] 678. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0772] 679. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0773] 680. The method of aspect 668, wherein said sulforaphane is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0774] 681. The method of aspect 622, wherein said histone deacetylase inhibitor is phenylbutyrate.

[0775] 682. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0776] 683. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0777] 684. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0778] 685. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0779] 686. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0780] 687. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0781] 688. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0782] 689. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0783] 690. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0784] 691. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0785] 692. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0786] 693. The method of aspect 668, wherein said phenylbutyrate is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0787] 694. The method of aspect 622, wherein said histone deacetylase inhibitor is entinostat.

[0788] 695. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0789] 696. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0790] 697. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0791] 698. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0792] 699. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0793] 700. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0794] 701. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0795] 702. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0796] 703. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0797] 704. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0798] 705. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0799] 706. The method of aspect 681, wherein said entinostat is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0800] 707. The method of aspect 622, wherein said histone deacetylase inhibitor is vorinostat.

[0801] 708. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of OCT4 more than 25%.

[0802] 709. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of OCT4 more than 50%.

[0803] 710. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of OCT4 more than 100%.

[0804] 711. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-kit more than 25%.

[0805] 712. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-kit more than 50%.

[0806] 713. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-kit more than 100%.

[0807] 714. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-met more than 25%.

[0808] 715. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-met more than 50%.

[0809] 716. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-met more than 100%.

[0810] 717. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-mpl more than 25%.

[0811] 718. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-mpl more than 50%.

[0812] 719. The method of aspect 707, wherein said vorinostat is administered at a concentration sufficient to induce expression of c-mpl more than 100%.

[0813] 720. The method of aspect 615, wherein said mesenchymal stem cell expressesCD73.

[0814] 721. The method of aspect 615, wherein said mesenchymal stem cell expressesCD105.

[0815] 722. The method of aspect 615, wherein said mesenchymal stem cell expresses c-kit.

[0816] 723. The method of aspect 615, wherein said mesenchymal stem cell expressesDAZZLE.

[0817] 724. The method of aspect 615, wherein said mesenchymal stem cell expressesHGF receptor.

[0818] 725. The method of aspect 616, wherein said mesenchymal stem cell expressesCD56.

[0819] 726. The method of aspect 616, wherein said mesenchymal stem cell expressesLIF receptor.

[0820] 727. The method of aspect 616, wherein said mesenchymal stem cell expressesCD90.

[0821] 728. The method of aspect 616, wherein said mesenchymal stem cell expressesPD-L1.

[0822] 729. The method of aspect 616, wherein said mesenchymal stem cell expresses activin receptor.

[0823] 730. The method of aspect 616, wherein said mesenchymal stem cell expresses endoglin receptor.

[0824] 731. The method of aspect 616, wherein said mesenchymal stem cell expressesFGF-1 receptor.

[0825] 732. The method of aspect 616, wherein said mesenchymal stem cell expresses interferon gamma receptor.

[0826] 733. The method of aspect 616, wherein said mesenchymal stem cell expressesTNF alpha receptor p55.

[0827] 734. The method of aspect 616, wherein said mesenchymal stem cell expressesTNF alpha receptor p75.

[0828] 735. The method of aspect 616, wherein said mesenchymal stem cell expressesNGF receptor.

[0829] 736. The method of aspect 616, wherein said mesenchymal stem cell expresses hCG receptor.

[0830] 737. The method of aspect 616, wherein said mesenchymal stem cell expresses progesterone receptor.

[0831] 738. The method of aspect 616, wherein said mesenchymal stem cell expresses testosterone receptor.

[0832] 739. The method of aspect 616, wherein said mesenchymal stem cell expressesKlotho receptor.

[0833] 740. The method of aspect 616, wherein said mesenchymal stem cell expresses interleukin 1 beta receptor.

[0834] 741. The method of aspect 616, wherein said mesenchymal stem cell expresses interleukin 7 receptor.

[0835] 742. The method of aspect 616, wherein said mesenchymal stem cell expresses interleukin 12 receptor.

[0836] 743. The method of aspect 616, wherein said mesenchymal stem cell expressesEGF receptor.

[0837] 744. The method of aspect 616, wherein said mesenchymal stem cell expresses angiopoietin receptor.

[0838] 745. The method of aspect 616, wherein said mesenchymal stem cell expressesG-CSF receptor.

[0839] 746. The method of aspect 616, wherein said mesenchymal stem cell expressesM-CSF receptor.

[0840] 747. The method of aspect 616, wherein said mesenchymal stem cell expressesGM-CSF receptor.

[0841] 748. The method of aspect 616, wherein said mesenchymal stem cell expressesTGF-beta receptor.

[0842] 749. The method of aspect 616, wherein said mesenchymal stem cell expresses endoglin receptor.

[0843] 750. The method of aspect 616, wherein said mesenchymal stem cell expresses calcitonin receptor.

[0844] 751. The method of aspect 506, wherein said cardiac specific progenitor cell expresses alpha actinin.

[0845] 752. The method of aspect 506, wherein said cardiac specific progenitor cell expresses troponin.

[0846] 753. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin.

[0847] 754. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin heavy chain.

[0848] 755. The method of aspect 506, wherein said cardiac specific progenitor cell expresses CD133.

[0849] 756. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin heavy chain and CD133.

[0850] 757. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2v.

[0851] 758. The method of aspect 506, wherein said cardiac specific progenitor cell expresses My20.

[0852] 759. The method of aspect 506, wherein said cardiac specific progenitor cell expresses cMHC NKX2-5.

[0853] 760. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MEF2c.

[0854] 761. The method of aspect 506, wherein said cardiac specific progenitor cell expresses STAT4.

[0855] 762. The method of aspect 506, wherein said cardiac specific progenitor cell expresses STAT6.

[0856] 763. The method of aspect 506, wherein said cardiac specific progenitor cell expresses My20 and CD133.

[0857] 764. The method of aspect 506, wherein said cardiac specific progenitor cell expresses My20 and CD34.

[0858] 765. The method of aspect 506, wherein said cardiac specific progenitor cell expresses My20 and c-met.

[0859] 766. The method of aspect 506, wherein said cardiac specific progenitor cell expresses My20 and c-kit.

[0860] 767. The method of aspect 506, wherein said cardiac specific progenitor cell expresses GATA4.

[0861] 768. The method of aspect 506, wherein said cardiac specific progenitor cell expresses ISLL cTNL.

[0862] 769. The method of aspect 506, wherein said cardiac specific progenitor cell expresses ISLL cTNl.

[0863] 770. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2a.

[0864] 771. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin heavy chain and CD133.

[0865] 772. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin heavy chain and CD34.

[0866] 773. The method of aspect 506, wherein said cardiac specific progenitor cell expresses myosin heavy chain and c-met.

[0867] 774. The method of aspect 506, wherein said cardiac specific progenitor cell expresses cMHC NKX2-5 and c-kit.

[0868] 775. The method of aspect 506, wherein said cardiac specific progenitor cell expresses cMHC NKX2-5 and CD133.

[0869] 776. The method of aspect 506, wherein said cardiac specific progenitor cell expresses cMHC NKX2-5 and CD34.

[0870] 777. The method of aspect 506, wherein said cardiac specific progenitor cell expresses cMHC NKX2-5 and c-met.

[0871] 778. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MEF2c and c-kit.

[0872] 779. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MEF2c and CD133.

[0873] 780. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MEF2c and CD34.

[0874] 781. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MEF2c and c-met.

[0875] 782. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2a and c-kit.

[0876] 783. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2a and CD133.

[0877] 784. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2a and CD34.

[0878] 785. The method of aspect 506, wherein said cardiac specific progenitor cell expresses MLC2a and c-met.

[0879] 786. The method of aspect 506, wherein differentiation of said pluripotent stem cell

[0880] 786. The method of aspect 506, wherein said pluripotent stem cell is treated with a WNT agonist.

[0881] 787. The method of aspect 786, wherein said WNT agonist is 1-azakenpaullone(9-Bromo-7,12-dihydro-pyrido[3',2':2,3]azepino[4,5-b]indol-6(5H)-one), BIO ((2'Z,3'E)-6- Bromoindirubin-3'-oxime).

[0882] 788. The method of aspect 786, wherein said WNT agonist is CHIR99021 (6-(2-(4-(2,4-dichlorophenyl)-5-(4-methyl-lH-imidazol-2-yl)pyrimidin-2- ylamino)ethylamino)nicotinonitrile).

[0883] 789. The method of aspect 786, wherein said WNT agonist is AR-A014418 (N-(4-Methoxybenzyl)-N'-(5-nitro-l,3-thiazol-2-yl)urea).

[0884] 790. The method of aspect 786, wherein said WNT agonist is lndirubin-3'- monoxime.

[0885] 791. The method of aspect 786, wherein said WNT agonist is 5-lodo-indirubin-3'-monoxime.

[0886] 792. The method of aspect 786, wherein said WNT agonist is kenpaullone (9-Bromo-7,12-dihydroindolo-[3,2-d][l]benzazepin-6(5H)-one).

[0887] 793. The method of aspect 786, wherein said WNT agonist is SB-415286 (3-[(3-Chloro-4-hydroxyphenyl)amino]-4-(2-nitro-phenyl)-lH-pyrrole-2, 5-dione).

[0888] 793. The method of aspect 786, wherein said WNT agonist is SB-216763 (3-(2,4-Dichlorophenyl)-4-(l-methyl-lH-indol-3-yl)-lH-pyrrole-2, 5-dione).

[0889] 794. The method of aspect 786, wherein said WNT agonist is MaybridgeSEW00923SC (2-anilino-5-phenyl-l,3,4-oxadiazole).

[0890] 795. The method of aspect 786, wherein said WNT agonist is (Z)-5-(2,3-Memylenedioxyphenyl)imidazolidine-2, 4-dione.

[0891] 796. The method of aspect 786, wherein said WNT agonist is TWS119 (3-(6-(3- aminophenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yloxy)phenol).

[0892] 797. The method of aspect 786, wherein said WNT agonist is CHIR98014 (N2-(2-(4-(2,4-dichlorophenyl)-5-(lH-imidazol-l-yl)pyrimidin-2-ylamino)ethyl)-5-nitropyridine-2,6-diamine).

[0893] 798. The method of aspect 786, wherein said WNT agonist is SB415286 (3-(3- chloro-4-hydroxyphenylamino)-4-(2-nitrophenyl)-lH-pyrrole-2, 5-dione).

[0894] 799. The method of aspect 786, wherein said WNT agonist is Tideglusib (also known as NP031112, or NP-12; 1, 2, 4-Thiadiazolidine-3, 5-dione, 2-(l-naphthalenyl)-4-(phenylmethyl)).

[0895] 800. The method of aspect 786, wherein said WNT agonist is LY2090314 (1H-Pyrrole-2, 5-dione, 3-imidazo[l,2-a]pyridin-3-yl-4-[l,2,3,4-tetrahydro-2-(l- piperidinylcarbonyl)pyrrolo[3,2,l-jk][l,4]benzodiazepin-7-yl]).

[0896] 801. The method of aspect 506, wherein said pluripotent stem cell is treated with a GSK-3 inhibitor.

[0897] 802. The method of aspect 801, wherein said GSK-3 inhibitor is lithium.

[0898] 803. The method of aspect 801, wherein said GSK-3 inhibitor is ChonglouSaponin VII.

[0899] 804. The method of aspect 801, wherein said GSK-3 inhibitor is MAZ51.

[0900] 805. The method of aspect 801, wherein said GSK-3 inhibitor is BRD0705.

[0901] 806. The method of aspect 801, wherein said GSK-3 inhibitor is Elragl usib (9-ING-41).

[0902] 807. The method of aspect 801, wherein said GSK-3 inhibitor is CP21.

[0903] 808. The method of aspect 801, wherein said GSK-3 inhibitor is BlO-acetoxime.

[0904] 809. The method of aspect 801, wherein said GSK-3 inhibitor is bikinin.

[0905] 810. The method of aspect 801, wherein said GSK-3 inhibitor is IM12.

[0906] 811. The method of aspect 801, wherein said GSK-3 inhibitor is AR-A014418.

[0907] 812. The method of aspect 802, wherein said lithium is administered together with valproic acid.

[0908] 813. The method of aspect 802, wherein said lithium is administered together with phenylbutyrate.

[0909] 814. The method of aspect 802, wherein said lithium is administered together with trichostatin A.

[0910] 815. The method of aspect 802, wherein said lithium is administered together with sulforaphane.

[0911] 816. The method of aspect 802, wherein said lithium is administered together with belinostat.

[0912] 817. The method of aspect 802, wherein said lithium is administered together with gavinostat.

[0913] 818. The method of aspect 802, wherein said lithium is administered together with interleukin-3.

[0914] 819. The method of aspect 802, wherein said lithium is administered together with interleukin-7.

[0915] 820. The method of aspect 802, wherein said lithium is administered together with interleukin-11.

[0916] 821. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with valproic acid.

[0917] 822. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with phenylbutyrate.

[0918] 823. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with trichostatin A.

[0919] 824. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with sulforaphane.

[0920] 825. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with belinostat.

[0921] 826. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with gavinostat.

[0922] 827. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with interleukin-3.

[0923] 828. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with interleukin-7.

[0924] 829. The method of aspect 803, wherein said Chonglou Saponin VII is administered together with interleukin-11.

[0925] 830. The method of aspect 804, wherein said MAZ51is administered together with valproic acid.

[0926] 831. The method of aspect 804, wherein said MAZ51 is administered together with phenylbutyrate.

[0927] 832. The method of aspect 804, wherein said MAZ51 is administered together with trichostatin A.

[0928] 833. The method of aspect 804, wherein said MAZ51 is administered together with sulforaphane.

[0929] 834. The method of aspect 804, wherein said MAZ51 is administered together with belinostat.

[0930] 835. The method of aspect 804, wherein said MAZ51 is administered together with gavinostat.

[0931] 836. The method of aspect 804, wherein said MAZ51 is administered together with interleukin-3.

[0932] 837. The method of aspect 804, wherein said MAZ51 is administered together with interleukin-7.

[0933] 838. The method of aspect 804, wherein said MAZ51 is administered together with interleukin-11.

[0934] 839. The method of aspect 805, wherein said BRD0705 is administered together with valproic acid.

[0935] 840. The method of aspect 805, wherein said BRD0705 is administered together with phenylbutyrate.

[0936] 841. The method of aspect 805, wherein said BRD0705 is administered together with trichostatin A.

[0937] 842. The method of aspect 805, wherein said BRD0705 is administered together with sulforaphane.

[0938] 843. The method of aspect 805, wherein said BRD0705 is administered together with belinostat.

[0939] 844. The method of aspect 805, wherein said BRD0705 is administered together with gavinostat.

[0940] 845. The method of aspect 805, wherein said BRD0705 is administered together with interleukin-3.

[0941] 846. The method of aspect 805, wherein said BRD0705 is administered together with interleukin-7.

[0942] 847. The method of aspect 805, wherein said BRD0705 is administered together with interleukin-11.

[0943] 848. The method of aspect 806, wherein said Elraglusib is administered together with valproic acid.

[0944] 849. The method of aspect 806, wherein said Elraglusib is administered together with phenylbutyrate.

[0945] 850. The method of aspect 806, wherein said Elraglusib is administered together with trichostatin A.

[0946] 851. The method of aspect 806, wherein said Elraglusib is administered together with sulforaphane.

[0947] 852. The method of aspect 806, wherein said Elraglusib is administered together with belinostat.

[0948] 853. The method of aspect 806, wherein said Elraglusib is administered together with gavinostat.

[0949] 854. The method of aspect 806, wherein said Elraglusib is administered together with interleukin-3.

[0950] 855. The method of aspect 806, wherein said Elraglusib is administered together with interleukin-7.

[0951] 856. The method of aspect 806, wherein said Elraglusib is administered together with interleukin-11.

[0952] 857. The method of aspect 807, wherein said CP21 is administered together with valproic acid.

[0953] 858. The method of aspect 807, wherein said CP21 is administered together with phenylbutyrate.

[0954] 859. The method of aspect 807, wherein said CP21 is administered together with trichostatin A.

[0955] 860. The method of aspect 807, wherein said CP21 is administered together with sulforaphane.

[0956] 861. The method of aspect 807, wherein said CP21 is administered together with belinostat.

[0957] 862. The method of aspect 807, wherein said CP21 is administered together with gavinostat.

[0958] 863. The method of aspect 807, wherein said CP21 is administered together with interleukin-3.

[0959] 864. The method of aspect 807, wherein said CP21 is administered together with interleukin-7.

[0960] 865. The method of aspect 807, wherein said CP21 is administered together with interleukin-11.

[0961] 866. The method of aspect 808, wherein said BlO-acetoxime is administered together with valproic acid.

[0962] 867. The method of aspect 808, wherein said BlO-acetoxime is administered together with phenylbutyrate.

[0963] 868. The method of aspect 808, wherein said BlO-acetoxime is administered together with trichostatin A.

[0964] 869. The method of aspect 808, wherein said BlO-acetoxime is administered together with sulforaphane.

[0965] 870. The method of aspect 808, wherein said BlO-acetoxime is administered together with belinostat.

[0966] 871. The method of aspect 808, wherein said BlO-acetoxime is administered together with gavinostat.

[0967] 872. The method of aspect 808, wherein said BlO-acetoxime is administered together with interleukin-3.

[0968] 873. The method of aspect 808, wherein said BlO-acetoxime is administered together with interleukin-7.

[0969] 874. The method of aspect 808, wherein said BlO-acetoxime is administered together with interleukin-11.

[0970] 875. The method of aspect 809, wherein said bikinin is administered together with valproic acid.

[0971] 876. The method of aspect 809, wherein said bikinin is administered together with phenylbutyrate.

[0972] 877. The method of aspect 809, wherein said bikinin is administered together with trichostatin A.

[0973] 878. The method of aspect 809, wherein said bikinin is administered together with sulforaphane.

[0974] 879. The method of aspect 809, wherein said bikinin is administered together with belinostat.

[0975] 880. The method of aspect 809, wherein said bikinin is administered together with gavinostat.

[0976] 881. The method of aspect 809, wherein said bikinin is administered together with interleukin-3.

[0977] 882. The method of aspect 809, wherein said bikinin is administered together with interleukin-7.

[0978] 883. The method of aspect 809, wherein said bikinin is administered together with interleukin-11.

[0979] 884. The method of aspect 810, wherein said IM12 is administered together with valproic acid.

[0980] 885. The method of aspect 810, wherein said IM12 is administered together with phenylbutyrate.

[0981] 886. The method of aspect 810, wherein said IM12 is administered together with trichostatin A.

[0982] 887. The method of aspect 810, wherein said IM12 is administered together with sulforaphane.

[0983] 888. The method of aspect 810, wherein said IM12 is administered together with belinostat.

[0984] 889. The method of aspect 810, wherein said IM12 is administered together with gavinostat.

[0985] 890. The method of aspect 810, wherein said IM12 is administered together with interleukin-3.

[0986] 891. The method of aspect 810, wherein said IM12 is administered together with interleukin-7.

[0987] 892. The method of aspect 810, wherein said IM12 is administered together with interleukin-11.

[0988] 893. The method of aspect 811, wherein said AR-A014418 is administered together with valproic acid.

[0989] 894. The method of aspect 811, wherein said AR-A014418 is administered together with phenylbutyrate.

[0990] 895. The method of aspect 811, wherein said AR-A014418 is administered together with trichostatin A.

[0991] 896. The method of aspect 811, wherein said AR-A014418 is administered together with sulforaphane.

[0992] 897. The method of aspect 811, wherein said AR-A014418 is administered together with belinostat.

[0993] 898. The method of aspect 811, wherein said AR-A014418 is administered together with gavinostat.

[0994] 899. The method of aspect 811, wherein said AR-A014418 is administered together with interleukin-3.

[0995] 900. The method of aspect 811, wherein said AR-A014418 is administered together with interleukin-7.

[0996] 901. The method of aspect 811, wherein said AR-A014418 is administered together with interleukin-11.

[0997] 902. A method of generating progenitor cells of a desired tissue from a pluripotent stem cell comprising the steps of: a) identifying embryonic cytokines associated with differentiation of said tissue; b) applying said cytokines to embryoid body derived from said pluripotent stem cells; and c) isolating and expanding said progenitor cells.

[0998] 903. The method of aspect 902, wherein said embryonically associated cytokines are detected by flow cytometry.

[0999] 904. The method of aspect 902, wherein said embryonically associated cytokines are detected by mass cytometry.

[1000] 905. The method of aspect 902, wherein said embryonically associated cytokines are detected by immunohistochemistry.

[1001] 906. The method of aspect 902, wherein said embryonically associated cytokines are detected by two photon microscopy.

[1002] 907. The method of aspect 902, wherein said embryonically associated cytokines are detected by GC-MS / MS.

[1003] 908. The method of aspect 902, wherein said embryonically associated cytokines are detected by laser capture microdissection.

[1004] 909. The method of aspect 902, wherein said embryonically associated cytokines are detected by microarray technology.

[1005] 910. The method of aspect 902, wherein said embryonically associated cytokines are detected by in situ hybridization.

[1006] 911. The method of aspect 902, inhibitors of differentiation are administered to allow for expansion of specific progenitor cells without substantial differentiation.

[1007] 912. The method of aspect 911, wherein said inhibitor of differentiation is a growth factor.

[1008] 913. The method of aspect 912, wherein said growth factor is a hematopoietic growth factor.

[1009] 914. The method of aspect 913, wherein said hematopoietic growth factor isEPO.

[1010] 915. The method of aspect 913, wherein said hematopoietic growth factor isGDNF.

[1011] 916. The method of aspect 913, wherein said hematopoietic growth factor is G-CSF.

[1012] 917. The method of aspect 913, wherein said hematopoietic growth factor isGM-CSF.

[1013] 918. The method of aspect 913, wherein said hematopoietic growth factor isGDF-9.

[1014] 919. The method of aspect 913, wherein said hematopoietic growth factor isHGF.

[1015] 920. The method of aspect 913, wherein said hematopoietic growth factor isHDGF.

[1016] 921. The method of aspect 913, wherein said hematopoietic growth factor isIGF.

[1017] 922. The method of aspect 913, wherein said hematopoietic growth factor isFas ligand immobilized to a matrix.

[1018] 923. The method of aspect 913, wherein said hematopoietic growth factor is migration stimulating factor.

[1019] 924. The method of aspect 913, wherein said hematopoietic growth factor isGDF-9.

[1020] 925. The method of aspect 913, wherein said hematopoietic growth factor isHGF.

[1021] 926. The method of aspect 913, wherein said hematopoietic growth factor isGDNF-8.

[1022] 927. The method of aspect 913, wherein said hematopoietic growth factor isGDNF-11.

[1023] 928. The method of aspect 913, wherein said hematopoietic growth factor isGDNF-15.

[1024] 929. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid.

[1025] 930. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with lithium.

[1026] 931. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with valproic acid.

[1027] 932. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with phenylbutyrate.

[1028] 933. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with sulforaphane.

[1029] 934. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with tricostatin A.

[1030] 935. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with monocyte conditioned media.

[1031] 936. The method of aspect 935, wherein said monocyte conditioned media is obtained by exposure of monocytes to allogeneic T cells.

[1032] 937. The method of aspect 936, wherein said allogeneic T cells express CD3.

[1033] 938. The method of aspect 936, wherein said allogeneic T cells express CD4.

[1034] 939. The method of aspect 936, wherein said allogeneic T cells express CD8.

[1035] 940. The method of aspect 936, wherein said allogeneic T cells express granzyme B.

[1036] 941. The method of aspect 936, wherein said allogeneic T cells express perforin.

[1037] 942. The method of aspect 936, wherein said allogeneic T cells express TNF- alpha upon ligation of CD3 and CD28.

[1038] 943. The method of aspect 936, wherein said allogeneic T cells express interferon alpha upon ligation of CD3 and CD28.

[1039] 944. The method of aspect 936, wherein said allogeneic T cells express lymphotoxin upon ligation of CD3 and CD28.

[1040] 945. The method of aspect 936, wherein said allogeneic T cells express interferon gamma upon ligation of CD3 and CD28.

[1041] 946. The method of aspect 936, wherein said allogeneic T cells express interleukin-1 beta upon ligation of CD3 and CD28.

[1042] 947. The method of aspect 936, wherein said allogeneic T cells express interleukin-17 upon ligation of CD3 and CD28.

[1043] 948. The method of aspect 936, wherein said allogeneic T cells express TRANCE upon ligation of CD3 and CD28.

[1044] 949. The method of aspect 936, wherein said allogeneic T cells express HLA II upon ligation of CD3 and CD28.

[1045] 950. The method of aspect 936, wherein said allogeneic T cells express transport associated protein 1 ligation of CD3 and CD28.

[1046] 951. The method of aspect 936, wherein said allogeneic T cells express TNF- alpha upon ligation of CD3 and CD28.

[1047] 952. The method of aspect 913, wherein said hematopoietic growth factor is hyaluronic acid mixed with mesenchymal stem cell conditioned media.

[1048] 953. The method of aspect 952, wherein said mesenchymal conditioned media is obtained by exposure of monocytes to allogeneic T cells.

[1049] 954. The method of aspect 953, wherein said allogeneic T cells express CD3.

[1050] 955. The method of aspect 953, wherein said allogeneic T cells express CD4.

[1051] 956. The method of aspect 953, wherein said allogeneic T cells express CD8.

[1052] 957. The method of aspect 953, wherein said allogeneic T cells express granzyme B.

[1053] 958. The method of aspect 953, wherein said allogeneic T cells express perforin.

[1054] 959. The method of aspect 953, wherein said allogeneic T cells express TNF- alpha upon ligation of CD3 and CD28.

[1055] 960. The method of aspect 953, wherein said allogeneic T cells express interferon alpha upon ligation of CD3 and CD28.

[1056] 961. The method of aspect 953, wherein said allogeneic T cells express lymphotoxin upon ligation of CD3 and CD28.

[1057] 962. The method of aspect 953, wherein said allogeneic T cells express interferon gamma upon ligation of CD3 and CD28.

[1058] 963. The method of aspect 953, wherein said allogeneic T cells express interleukin-1 beta upon ligation of CD3 and CD28.

[1059] 964. The method of aspect 953, wherein said allogeneic T cells express interleukin-17 upon ligation of CD3 and CD28.

[1060] 965. The method of aspect 953, wherein said allogeneic T cells express TRANCE upon ligation of CD3 and CD28.

[1061] 966. The method of aspect 953, wherein said mesenchymal stem cells are pretreated with an activator of NF-kappa B.

[1062] 967. The method of aspect 966, wherein said activator of NF-kappa B is an activator of a toll like receptor.

[1063] 968. The method of aspect 966, wherein said activator of NF-kappa B is an activator of MDA.

[1064] 969. The method of aspect 966, wherein said activator of NF-kappa B is an activator of RIG-1.

[1065] 970. The method of aspect 966, wherein said activator of NF-kappa B is an activator of MAP kinase.

[1066] 971. The method of aspect 966, wherein said NF-kappa B activator is quercetin.

[1067] 972. The method of aspect 966, wherein said NF-kappa B activator is DNA extracellular traps

[1068] 973. The method of aspect 966, wherein said NF-kappa B activator is zymosan.

[1069] 974. The method of aspect 966, wherein said NF-kappa B activator is HMGB1.

[1070] 975. The method of aspect 966, wherein said NF-kappa B activator is TNF-alpha.

[1071] 976. The method of aspect 966, wherein said NF-kappa B activator is crosslinking antibody to TLR3.

[1072] 977. The method of aspect 966, wherein said NF-kappa B activator is crosslinking antibody to TLR4.

[1073] 978. The method of aspect 966, wherein said NF-kappa B activator is crosslinking antibody to TLR7.

[1074] 979. The method of aspect 966, wherein said NF-kappa B activator is crosslinking antibody to TLR9.

[1075] 980. The method of aspect 966, wherein said NF-kappa B activator is imiquimod.

[1076] 981. The method of aspect 966, wherein said NF-kappa B activator is resiminod.

[1077] 982. The method of aspect 966, wherein said NF-kappa B activator is lipopolysaccharide.

[1078] 983. The method of aspect 966, wherein said NF-kappa B activator is beta glucan.

[1079] 984. The method of aspect 966, wherein said NF-kappa B activator is CpG DNA motifs.

[1080] 985. The method of aspect 966, wherein said NF-kappa B activator is double stranded RNA.

[1081] 986. The method of aspect 966, wherein said NF-kappa B activator is circularDNA.

[1082] 987. The method of aspect 966, wherein said NF-kappa B activator is uric acid crystals.

[1083] 988. The method of aspect 966, wherein said NF-kappa B activator is yeast cell wall extract.

[1084] 989. The method of aspect 966, wherein said NF-kappa B activator is immunomax.

[1085] 990. The method of aspect 966, wherein said NF-kappa B activator is activate neutrophils.

[1086] 991. The method of aspect 966, wherein said NF-kappa B activator is interleukin-6.

[1087] 992. The method of aspect 966, wherein said NF-kappa B activator is interleukin-8.

[1088] 993. The method of aspect 966, wherein said NF-kappa B activator is interleukin-11.

[1089] 994. The method of aspect 966, wherein said NF-kappa B activator is interleukin-12.

[1090] 995. The method of aspect 966, wherein said NF-kappa B activator is interleukin-15.

[1091] 996. The method of aspect 966, wherein said NF-kappa B activator is interleukin-17.

[1092] 997. The method of aspect 966, wherein said NF-kappa B activator is interleukin-18.

[1093] 998. The method of aspect 966, wherein said NF-kappa B activator is interleukin-23.

[1094] 999. The method of aspect 966, wherein said NF-kappa B activator is interleukin-27.

[1095] 1000. The method of aspect 966, wherein said NF-kappa B activator is interleukin-33.

[1096] 1001. The method of aspect 912, wherein said growth factor is selected from a group of growth factors comprising of: AM, Ang, BMP, BDNF, EGF, FGF, or a Wnt protein; an interleukin; a soluble receptor for IL-1. alpha., IL-1. beta., IL-1F1, IL-1F2, IL-1F3, IL-1F4, IL-1F5, IL-1F6, IL-1F7, IL-1F8, IL-1F9, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-1235 kDa alpha subunit, IL-1240 kDa beta subunit, IL-13, IL-14, IL-15, IL-16, IL-17A, IL- 176, IL-17C, IL-17D, IL-17E, IL-17F isoform 1, IL-17F isoform 2, IL-18, IL-19, IL-20, IL-21, IL- 22, IL-23 pl9 subunit, IL-23 p40 subunit, IL-24, IL-25, IL-26, IL-27B, IL-27-p28, IL-28A, IL- 286, IL-29, IL-30, IL-31, IL-32, IL-33, IL-34, IL-35, IL-36.alpha., IL-36.beta., IL-36.gamma.;an interferon (IFN); a soluble receptor for IFN-.alpha., IFN-.beta., IFN-.gamma., IFN- .lamda.l, IFN-.lamda.2, IFN-.lamda.3, IFN-K, IFN-.epsilon., IFN-.kappa., IFN-.tau., IFN- . delta., IFN-.zeta., IFN-.omega., or IFN-v; insulin or proinsulin; a receptor for insulin; leptin (LEP).BRIEF DESCRI PTION OF DRAWINGS

[1097] Figure 1 is a bar graph showing the results of alpha myosin heavy chain expression in generated progenitor cells in relation to whether conditioned media, valproic acid, or conditioned media + valproic acid was used.

[1098] Figure 2 is a bar graph showing the results of c-kit expression in generated progenitor cells in relation to whether conditioned media, valproic acid, or conditioned media + valproic acid was used.

[1099] Figure 3 is a bar graph showing the results of CD133 expression in generated progenitor cells in relation to whether conditioned media, valproic acid, or conditioned media + valproic acid was used.DETAILED DESCRIPTION OF TH E I NVENTION

[1100] The invention provides means of generating tissue specific progenitor cells from pluripotent stem cells. In one embodiment the invention provides means of differentiating pluripotent stem cells into tissue specific cells by culturing of said pluripotent stem cells with factors released from tissue or cell lines representing tissue, of the type of which differentiated progenitor cells are desired. In one embodiment the invention teaches the creation of cell lines, for example, cardiac cell lines, which are treated with stressors, and factors produced by said cell lines are utilized to induce pluripotent stem cell differentiation. In other embodiments the invention provides the administration of specific growth factors that replicate embryonic development of tissue whose production is desired. In other embodiments microvesicles, exosomes, microRNAs and circular DNA

[1101] As used herein, "Cardiomyocytes" or cardiac myocytes are the muscle cells that make up the cardiac muscle. Each myocardial cell contains myofibrils, which are long chains of sarcomeres, the contractile units of muscle cells. Cardiomyocytes show striations similar to those on skeletal muscle cells, but unlike multinucleated skeletalcells, they contain only one nucleus. Cardiomyocytes have a high mitochondrial density, which allows them to produce ATP quickly, making them highly resistant to fatigue. Mature cardiomyocytes can express one or more of the following cardiac markers: a- Actinin, MLC2v, MY20, cMHC, NKX2-5, GATA4, cTNT, cTNI, MEF2c, MLC2a, or any combination thereof. In some embodiments, the mature cardiomyocytes express NKX2- 5, MEF2c or a combination thereof. Cardiac progenitor cells express early stage cardiac progenitor markers such as GATA4, ISL1 or a combination thereof.

[1102] As used herein, the term "functional cardiomyocyte" refers to a differentiated cardiomyocyte that is able to send or receive electrical signals. In some embodiments, a cardiomyocyte is said to be a functional cardiomyocyte if it exhibits electrophysiological properties such as action potentials and / or Ca2+ transients.

[1103] As used herein, a "differentiated non-cardiac cell" can refer to a cell that is not able to differentiate into all cell types of an adult organism (i.e., is not a pluripotent cell), and which is of a cellular lineage other than a cardiac lineage (e.g., fibroblast, a cell of endodermal, mesodermal, epithelial, neuronal, connective, lymphocyte, or other tissue type lineage). Differentiated cells include, but are not limited to, multipotent cells, oligopotent cells, unipotent cells, progenitor cells, and terminally differentiated cells. In particular embodiments, a less potent cell is considered "differentiated" in reference to a more potent cell.

[1104] As used herein, a cell that differentiates into a mesodermal, ectodermal or endodermal lineage defines a cell that becomes committed to a specific mesodermal, ectodermal or endodermal lineage, respectively. Examples of cells that differentiate into a mesodermal lineage or give rise to specific mesodermal cells include, but are not limited to, cells that are adipogenic, chondrogenic, cardiogenic, dermatogenic, hematopoietic, hemangiogenic, myogenic, nephrogenic, urogenitogenic, osteogenic, pericardiogenic, or stromal. Examples of cells that differentiate into ectodermal lineage include, but are not limited to epidermal cells, neurogenic cells, and neurogliagenic cells. Examples of cells that differentiate into endodermal lineage include, but are not limited to pleurigenic cells, and hepatogenic cells, that give rise to the lining of the intestine, and cells that give rise to pancreogenic and splanchogenic cells.

[1105] As used herein, the term "cell culture medium" (also referred to herein as a "culture medium" or "medium") as referred to herein is a medium for culturing cells containing nutrients that maintain cell viability and support proliferation. The cell culture medium can contain any of the following in an appropriate combination: salt(s),buffer(s), amino acids, glucose or other sugar(s), antibiotics, serum or serum replacement, and other components such as peptide growth factors, etc. Cell culture media ordinarily used for particular cell types are available to those skilled in the art.

[1106] A "somatic cell" is a cell forming the body of an organism. Somatic cells include cells making up organs, skin, blood, bones and connective tissue in an organism, but not germ cells.

[1107] Cells can be from, e.g., human or non-human mammals. Exemplary non-human mammals include, but are not limited to, mice, rats, cats, dogs, rabbits, guinea pigs, hamsters, sheep, pigs, horses, bovines, and non-human primates. In some embodiments, a cell is from an adult human or non-human mammal. In some embodiments, a cell is from a neonatal human, an adult human, or non-human mammal.

[1108] As used herein, the term "totipotent" means the ability of a cell to form all cell lineages of an organism. For example, in mammals, only the zygote and the first cleavage stage blastomeres are totipotent.

[1109] As used herein, the term "pluripotent" means the ability of a cell to form all lineages of the body or soma (i.e., the embryo proper). For example, embryonic stem cells are a type of pluripotent stem cells that are able to form cells from each of the three germs layers, the ectoderm, the mesoderm, and the endoderm.

[1110] As used herein, the term "multipotent" refers to the ability of an adult stem cell to form multiple cell types of one lineage. For example, hematopoietic stem cells are capable of forming all cells of the blood cell lineage, e.g., lymphoid and myeloid cells.

[1111] As used herein, the term "oligopotent" refers to the ability of an adult stem cell to differentiate into only a few different cell types. For example, lymphoid or myeloid stem cells are capable of forming cells of either the lymphoid or myeloid lineages, respectively.

[1112] As used herein, the term "unipotent" means the ability of a cell to form a single cell type. For example, spermatogonial stem cells are only capable of forming sperm cells.

[1113] As used herein, the term "direct reprogramming" or "transdifferentiation" refers to the generation of a cell of a certain lineage (e.g., a cardiac cell) from a different type of cell (e.g., a fibroblast cell) without an intermediate process of de-differentiating the cell into a cell exhibiting pluripotent stem cell characteristics.

[1114] As used herein, the terms "subject" or "patient" refers to any animal, such as a domesticated animal, a zoo animal, or a human. The "subject" or "patient" can be a mammal like a dog, cat, bird, livestock, or a human. Specific examples of "subjects" and "patients" include, but are not limited to, individuals with a cardiac disease or disorder, and individuals with cardiac disorder-related characteristics or symptoms.

[1115] In one embodiment of the invention, pluripotent stem cells are differentiated into progenitors of various tissues by using various means of tissue specific differentiation based on embryologically expressed factors associated with differentiation. Depending on the tissue various factors can be administered. Said factors include, bone morphogenic protein (BMP)-l, bone morphogenic protein-2, bone morphogenic protein-3, bone morphogenic protein-4, bone morphogenic protein-5, bone morphogenic protein-6, bone morphogenic protein-7, bone morphogenic protein- 8, bone morphogenic protein-9, bone morphogenic protein-10, bone morphogenic protein-11, bone morphogenic protein-12, bone morphogenic protein-13, bone morphogenic protein-14, bone morphogenic protein-15, brain derived neurotrophic factor, ciliary neurotrophic factor, cytokine-induced neutrophil chemotactic factor 1, cytokine-induced neutrophil chemotactic factor 2a, cytokine-induced neutrophil chemotactic factor 2P, p endothelial cell growth factor, endothelin 1, epidermal growth factor, epithelial-derived neutrophil attractant, fibroblast growth factor (FGF) 4, fibroblast growth factor 5, fibroblast growth factor 6, fibroblast growth factor 7, fibroblast growth factor 8, fibroblast growth factor 8b, fibroblast growth factor 8c, fibroblast growth factor 9, fibroblast growth factor 10, fibroblast growth factor (acidic), fibroblast growth factor (basic), growth related protein, growth related protein a, growth related protein p, growth related protein y, heparin binding epidermal growth factor, hepatocyte growth factor, insulin-like growth factor I, insulin-like growth factor II, insulin-like growth factor binding protein, keratinocyte growth factor, leukemia inhibitory factor, neurotrophin-3, neurotrophin-4, placenta growth factor, placenta growth factor 2, platelet-derived endothelial cell growth factor, platelet derived growth factor, platelet derived growth factor A chain, platelet derived growth factor AA, platelet derived growth factor AB, platelet derived growth factor B chain, platelet derived growth factor BB, pre-B cell growthstimulating factor, stem cell factor, transforming growth factor a, transforming growth factor p, transforming growth factor pi, transforming growth factor pi.2, transforming growth factor P2, transforming growth factor P3, latent transforming growth factor pi, transforming growth factor p bindingprotein I, transforming growth factor p binding protein 11, transforming growth factor binding protein 111, and vascular endothelial growth factor. Exemplary cytokines can be included such as interleukin (IL)-l, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL- 12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, interferon (IFN), IFN-y, tumor necrosis factor (TNF), TNF1, TNF2, TNF-a, macrophage colony stimulating factor (M-CSF), granulocytemonocyte colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G- CSF), megakaryocyte colony stimulating factor (Meg-CSF)-thrombopoietin, stem cell factor, and erythropoietin. Chemokines can also be included such as IP-10 and Stromal Cell-Derived Factor la. While the previously mentioned factors possess various differentiation inducing ability, specific cellular types can be generated using more specific productions means.

[1116] For example, in situations in which cardiac progenitor cells are desired, cells can be incubated with a reprogramming composition that contains one or more GSK3 inhibitors / WNT agonists, TGF-beta inhibitors, inhibitors of extracellular signal-regulated kinase 1 (ERK1), inhibitors of Ras GTPase-activating protein (Ras-GAP)), Oct-4 activators, P160ROCK inhibitors (where pl60ROCK is a rho-associated protein kinase), iron chelator and / or KDM5B inhibitor, inhibitors of G9a histone methyltransferase, inhibitors of various growth factor receptors such as PDGF receptor beta, protein kinase receptor inhibitors, inhibitors of PDGF-BB receptor, and any combination thereof. The composition can contain at least two of the agents, or at least three of the agents, or at least four of the agents, or at least five of the agents, or at least six of the agents, or at least seven of the agents, or at least eight of the agents. After incubation in a reprogramming medium, the cells can then incubated in the first and / or second cardiac induction media. The base media employed to which the reprogramming agents or induction agents are added can be a convenient cell culture medium. Examples cell culture media that can be employed include mTESR-1® medium (StemCell Technologies, Inc., Vancouver, Calif.), or Essential 8® medium (Life Technologies, Inc.) on a Matrigel substrate (BD Biosciences, NJ) or on a Corning® Synthemax surface, or in Johansson and Wiles CDM supplemented with insulin, transferrin, lipids and polyvinyl alcohol (PVA) as substitute for Bovine Serum Albumin (BSA). Examples of commercially available media also include, but are not limited to, Dulbecco's Modified Eagle's Medium (DMEM), Minimal Essential Medium (MEM), Basal Medium Eagle (BME), RPMI 1640, Ham's F-10, Ham's F-12, a-Minimal Essential Medium (aMEM), Glasgow's Minimal Essential Medium(G-MEM), Iscove's Modified Dulbecco's Medium, or a general purpose media modified for use with pluripotent cells, such as X-VIVO (Lonza) or a hematopoietic base media.

[1117] The starting cells can be dispersed in a cell culture medium that contains the reprogramming composition at a density that permits cell expansion. For example, about 1 to 1010 cells can be contacted with the reprogramming composition in a selected cell culture medium, especially when the cells are maintained at a cell density of about 1 to about 108 cells per milliliter, or at a density of about 100 to about 107 cells per milliliter, or at a density of about 1000 to about 106 cells per milliliter.

[1118] The time for conversion of starting cells into cardiac progenitor and cardiomyocyte cells can vary. For example, the starting cells can be incubated with the reprogramming composition until cardiac or cardiomyocyte cell markers are expressed. Such cardiac or cardiomyocyte cell markers can include any of the following markers: a- Actinin, MLC2v, MY20, cMHC, NKX2-5, MEF2c, GATA4, ISL1, cTNT, cTNI, MLC2a and any combination thereof.

[1119] Incubation can proceed in any of the compositions described herein, for example, until early stage cardiac progenitor markers are expressed by the starting cells. Such early stage cardiac progenitor markers include GATA4, ISL1 or a combination thereof. The early stage cardiac progenitor markers such as GATA4 and / or ISL1 can be expressed by about 6 days, or by about 8 days, or by about 9 days, or by about 10 days, or by about 11 days, or by about 12 days of incubation of cells using the compositions and methods described herein.

[1120] Further incubation of the cells can be performed until expression of late stage cardiac progenitor markers such as NKX2-5, MEF2c or a combination thereof occurs. The late stage cardiac progenitor marker such as NKX2-5 and / or MEF2c can be expressed by about 14 days, or by about 15 days, or by about 16 days, or by about 17, or by about 18 days of incubation of cells using the compositions and methods described herein.

[1121] In some embodiments, the starting cells can be incubated with the reprogramming medium under cell culture conditions for about 1 day to about 15 days, or about 2 days to about 12 days, or about 3 days to about 10 days, or about 4 days to about 8 days, or about 5 days to about 7 days, or about 6 days.

[1122] After incubation in the reprogramming medium, maturation of the cardiac phenotype can be induced by incubation of the cells in a first cardiac induction medium (e.g., CIM1) for about 1 day to about 20 days, or about 2 days to about 14 days, or about 3 days to about 8 days, or about 4 days to about 6 days, or about 5 days.

[1123] After incubation in such a first cardiac induction medium (e.g., CIM1), the cells can be replated into a second cardiac induction medium (e.g., CIM2), and incubated for about 1 day to about 40 days, or about 3 days to about 36 days, or about 5 days to about 33 days, or about 7 days to about 30 days, or about 10 days to about 27 days, or about 15 days to about 25 days, or about 20 to about 22 days.

[1124] Cardiomyocytes exhibit some cardiac-specific electrophysiological properties.One electrical / physiological characteristic is an action potential, which is a short-lasting event in which the difference of potential between the interior and the exterior of each cardiac cell rises and falls following a consistent trajectory. Another electrophysiological characteristic of cardiomyocytes is the cyclic variations in the cytosolic-free Ca2+ concentration, named as Ca2+ transients, which are employed in the regulation of the contraction and relaxation of cardiomyocytes. These characteristics can be detected and evaluated to assess whether a population of cells has been reprogrammed into cardiomyocytes.

[1125] Such methods can therefore be used to generate a population of cardiac progenitor cells or cardiomyocytes that can be transplanted into a subject or used for experimentation.

[1126] In some embodiments, a reprogrammed population of cells (at various stages of reprogramming) can be frozen at liquid nitrogen temperatures, stored for periods of time, and then thawed for use at a later date. If frozen, a population of reprogrammed cells can be stored in a 10% DMSO, 50% FCS, within 40% RPMI 1640 medium. Once thawed, the cells can be expanded by culturing the cells in an appropriate medium that can contain selected growth factors, vitamins, feeder cells, and other components selected by a person of skill in the art.

[1127] Reprogrammed cells generated as described herein can be employed for tissue reconstitution or regeneration in a human patient or other subjects in need of such treatment. The cells are administered in a manner that permits them to graft or migrate to a diseased or injured tissue site and to reconstitute or regenerate the functionally deficient area. Devices are available that can be adapted for administering cells, for example, to cardiac tissues.

[1128] For therapy, reprogrammed cardiac progenitor cells, cardiomyocytes and / or pharmaceutical compositions can be administered locally or systemically. A population of reprogrammed cells can be introduced by injection, catheter, implantable device, or the like. A population of reprogrammed cells can be administered in any physiologicallyacceptable excipient or carrier that does not adversely affect the cells. For example, the cardiac progenitor cells, cardiomyocytes and / or pharmaceutical compositions can be administered intravenously or through an intracardiac route (e.g., epicardial ly or intramyocardially). Methods of administering the cardiac progenitor cells, cardiomyocytes and / or pharmaceutical compositions of the invention to subjects, particularly human subjects, include injection or implantation of the cells into target sites in the subjects. The cells of the invention can be inserted into a delivery device which facilitates introduction of the cells after injection or implantation of the device within subjects. Such delivery devices include tubes, e.g., catheters, for injecting cells and fluids into the body of a recipient subject. The tubes can additionally include a needle, e.g., a syringe, through which the cells of the invention can be introduced into the subject at a desired location. The kits described herein can include such devices.

[1129] The cardiac progenitor cells and cardiomyocytes can be inserted into such a delivery device, e.g., a syringe, in different forms. A population of reprogrammed cells can be supplied in the form of a pharmaceutical composition. Such a composition can include an isotonic excipient prepared under sufficiently sterile conditions for human administration. For general principles in medicinal formulation, the reader is referred to CELL THERAPY: STEM CELL TRANSPLANTATION, GENE THERAPY, AND CELLULAR IMMUNOTHERAPY, by G. Morstyn & W. Sheridan eds, Cambridge University Press, 1996; and HEMATOPOIETIC STEM CELL THERAPY, E. D. Ball, J. Lister & P. Law, Churchill Livingstone, 2000. The choice of the cellular excipient and any accompanying constituents of the composition that includes a population of reprogrammed cells can be adapted to optimize administration by the route and / or device employed.

[1130] As used herein, the term "solution" includes a carrier or diluent in which the cardiomyocytes of the invention remain viable. Carriers and diluents which can be used with this aspect of the invention include saline, aqueous buffer solutions, physiologically acceptable solvents, and / or dispersion media. The use of such carriers and diluents is well known in the art. The solution is preferably sterile and fluid to allow syringability. For transplantation, a solution containing a suspension of cardiomyocytes can be drawn up into a syringe, and the solution containing the cells can be administrated to anesthetized transplantation recipients. Multiple injections may be made using this procedure.

[1131] The compositions, cardiac progenitor cells and / or cardiomyocytes can also be embedded in a support matrix. A composition that includes a population ofreprogrammed cells can also include or be accompanied by one or more other ingredients that facilitate engraftment or functional mobilization of the reprogrammed cells. Suitable ingredients include matrix proteins that support or promote adhesion of the reprogrammed cells, or complementary cell types, such cardiac pacemaker cells, or cardiac cells at different stages of maturation. In another embodiment, the composition may include physiologically acceptable matrix scaffolds. Such physiologically acceptable matrix scaffolds can be resorbable and / or biodegradable.

[1132] The population of reprogrammed cells generated by the methods described herein can include low percentages of non-cardiac cells (e.g., fibroblasts). For example, a population of reprogrammed cells for use in compositions and for administration to subjects can have less than about 90% non-cardiac cells, less than about 85% noncardiac cells, less than about 80% non-cardiac cells, less than about 75% non-cardiac cells, less than about 70% non-cardiac cells, less than about 65% non-cardiac cells, less than about 60% non-cardiac cells, less than about 55% non-cardiac cells, less than about 50% non-cardiac cells, less than about 45% non-cardiac cells, less than about 40% noncardiac cells, less than about 35% non-cardiac cells, less than about 30% non-cardiac cells, less than about 25% non-cardiac cells, less than about 20% non-cardiac cells, less than about 15% non-cardiac cells, less than about 12% non-cardiac cells, less than about 10% non-cardiac cells, less than about 8% non-cardiac cells, less than about 6% noncardiac cells, less than about 5% non-cardiac cells, less than about 4% non-cardiac cells, less than about 3% non-cardiac cells, less than about 2% non-cardiac cells, or less than about 1% non-cardiac cells of the total cells in the cell population.

[1133] Many cell types are capable of migrating to an appropriate site for regeneration and differentiation within a subject. To determine the suitability of various therapeutic administration regimens and dosages of cell compositions, the cells can first be tested in a suitable animal model. At one level, cells are assessed for their ability to survive and maintain their phenotype in vive. Cells can also be assessed to ascertain whether they migrate to diseased or injured sites in vivo, or to determine an appropriate dosage such as an appropriate number of cells and / or a frequency of administration of cells. Cell compositions can be administered to immunodeficient animals (such as nude mice, or animals rendered immunodeficient chemically or by irradiation). Tissues can be harvested after a period of regrowth, and assessed as to whether the administered cells or progeny thereof are still present, are alive, and / or have migrated to desired or undesired locations.

[1134] Injected cells can be traced by a variety of methods. For example, cells containing or expressing a detectable label (such as green fluorescent protein, or betagalactosidase) can readily be detected. The cells can be pre-labeled, for example, with BrdU or [3H]-thymidine, or by introduction of an expression cassette that can express green fluorescent protein, or beta-galactosidase. Alternatively, the reprogrammed cells can be detected by their expression of a cell marker that is not expressed by the animal employed for testing (for example, a human-specific antigen). The presence and phenotype of the administered population of reprogrammed cells can be assessed by fluorescence microscopy (e.g., for green fluorescent protein, or beta-galactosidase), by immunohistochemistry (e.g., using an antibody against a human antigen), by ELISA (using an antibody against a human antigen), or by RT-PCR analysis using primers and hybridization conditions that cause amplification to be specific for human polynucleotides.

[1135] The dose and the number of administrations can therefore be optimized by those skilled in the art.Pharmaceutical Compositions

[1136] The invention also relates to reprogramming compositions containing one or more of the following chemical agents: a GSK3 inhibitor, a WNT agonist, a TGF-beta inhibitor, an inhibitor of extracellular signal-regulated kinase 1 (ERK1), an inhibitor of Ras GTPase-activating protein (Ras-GAP), an Oct -4 activator, a pl60ROCK inhibitor (where P160ROCK is a rho-associated protein kinase), an iron chelator and / or KDM5B inhibitor, an inhibitor of G9a histone methyltransferase, an inhibitor of various growth factor receptors such as PDGF receptor beta, a protein kinase receptor inhibitor, an inhibitor of PDGF-BB receptor, and any combination thereof. For example, the composition can contain at least two of the agents, or at least three of the agents, or at least four of the agents, or at least five of the agents, or at least six of the agents, or at least seven of the agents, or at least eight of the agents, or at least nine of the agents. The compositions can also contain reprogrammed cells.

[1137] For example, such a reprogramming composition (9C) can include CHIR99021, A83-01, SCI, OAC, Y27632, BIX-01294, AS8351, SU16f, and JNJ-10198409. As described herein PBIT can be employed instead of AS8351.

[1138] The invention also relates to separate cardiac induction compositions containing one or more of the following chemical agents: a glycogen synthase kinase 3 (GSK3)inhibitor, and growth factors such as a bone morphogenic protein, a member of the TGFP family, vascular endothelial growth factor (VEGF), or any combination thereof.

[1139] The compositions of the invention can be pharmaceutical compositions. In some embodiments, the compositions can include a pharmaceutically acceptable carrier. By "pharmaceutically acceptable" it is meant a carrier, diluent, excipient, and / or salt that is compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof.

[1140] The compositions can contain any of the agent(s) or compound(s) described herein in an amount sufficient to reprogram a cell into a cardiac cell type. For example, the compositions can contain any of the agent(s) or compound(s) described herein in an amount sufficient to induce a cell to express cardiac or cardiomyocyte cell markers. Such cardiac or cardiomyocyte cell markers can include any of the following markers: a- Actinin, MLC2v, MY20, cMHC, NKX2-5, MEF2c, GATA4, ISL1, cTNT, cTNI, MLC2a and any combination thereof. Incubation can proceed in any of the compositions described herein, for example, until early stage cardiac progenitor markers are expressed by the starting cells. Such early stage cardiac progenitor markers include GATA4. ISL1 or a combination thereof. The early stage cardiac progenitor markers such as GATA4 and / or ISL1 can be expressed by about 6 days, or by about 8 days, or by about 9 days, or by about 10 days, or by about 11 days, or by about 12 days of incubation of cells using the compositions and methods described herein.

[1141] In some embodiments, the therapeutic compositions are administered in a "therapeutically effective amount." Such a therapeutically effective amount is an amount sufficient to obtain the desired physiological effect, e.g., treatment of a condition, disorder, disease and the like or reduction in symptoms of the condition, disorder, disease and the like. For example, the therapeutic agents can be administered to treat any of the conditions, disorders, or diseases described herein. Examples include congestive heart failure, myocardial infarction, cardiac ischemia myocarditis, arrhythmia or any combination thereof.

[1142] To achieve the desired effect(s), the composition can be formulated in single or divided dosages. For example, a GSK3 inhibitor, a WNT agonist, a TGF-beta inhibitor, an inhibitor of extracellular signal-regulated kinase 1 (ERK1), an inhibitor of Ras GTPase- activating protein (Ras-GAP), an Oct -4 activator, a pl60ROCK inhibitor (where pl60ROCK is a rho-associated protein kinase), an iron chelator and / or KDM5B inhibitor, an inhibitor of G9a histone methyltransferase, an inhibitor of various growth factor receptors such asPDGF receptor beta, a protein kinase receptor inhibitor, an inhibitor of PDGF-BB receptor, and / or a growth factor (e.g., any of the CIMl growth factors) can present in the composition in amounts specified above or in dosages of at least about 0.01 mg / kg to about 500 to 750 mg / kg, of at least about 0.01 mg / kg to about 300 to 500 mg / kg, at least about 0.1 mg / kg to about 100 to 300 mg / kg or at least about 1 mg / kg to about 50 to 100 mg / kg of body weight, although other dosages may provide beneficial results. The amount administered will vary depending on various factors including, but not limited to the combination of compounds chosen for administration, the disease, the weight, the physical condition, the health, the age of the mammal, as well as other physiological factors. Such factors can be readily determined by the clinician employing animal models or other test systems that are available in the art.

[1143] For example, a reprogramming composition can include CHIR99021 (e.g., at about 10-20 pM). A83-01 (e.g., at about 0.5-1.0 pM), SCI (e.g., at about 0.5-1.0 pM), OAC (e.g., at about 1-10 pM), Y27632 (e.g., at about 5-10 pM), BIX-01294 (e.g., at about 0.5-2.0 pM), AS8351 (e.g., at about 1.0-3.0 pM), SU16f (e.g., at about 2-5 pM), and JNJ- 10198409 (e.g., at about 0.05-0.2 pM). As described herein PBIT (e.g., at about 10 pM).

[1144] Reprogrammed cells can be included in the compositions in varying amounts depending upon the disease or injury to be treated. For example, the compositions can be prepared in liquid form for local or systemic administration containing about 103 to about 1012 reprogrammed cells, or about 104 to about 1010 reprogrammed cells, or about 105 to about 108 reprogrammed cells. One or more of the following types of compounds can also be present in the composition with the cells: a GSK3 inhibitor, a WNT agonist, a TGF-beta inhibitor, an inhibitor of extracellular signal-regulated kinase 1 (ERK1), an inhibitor of Ras GTPase-activating protein (Ras-GAP), an Oct-4 activator, a P160ROCK inhibitor (where pl60ROCK is a rho-associated protein kinase), an iron chelator and / or KDM5B inhibitor, an inhibitor of G9a histone methyltransferase, an inhibitor of various growth factor receptors such as PDGF receptor beta, a protein kinase receptor inhibitor, an inhibitor of PDGF-BB receptor, and / or one or more growth factors (e.g., any of the CIMl growth factors).

[1145] Administration of the composition may be in a single dose, in multiple doses, in a continuous or intermittent manner, depending, for example, upon the recipient's physiological condition, whether the purpose of the administration is for response to traumatic injury or for more sustained therapeutic purposes, and other factors known to skilled practitioners. Similarly, cell(s) can be contacted with the composition in acontinuous manner, or intermittently, depending upon the need for reprogrammed cells, the manufacturing schedule, the convenience of workers, and / or the selected recipient's physiological condition. The administration or contacting of the cells with compositions of the invention may be essentially continuous over a preselected period of time or may be in a series of spaced doses. Both local and systemic administration to recipients and / or subjects is contemplated.

[1146] To prepare the composition, the compounds are synthesized and / or the cells are generated, and the components are purified as necessary or desired. The compounds, cells, and / or other agents can be suspended in a pharmaceutically acceptable carrier. If the composition contains only compounds, without cells, the composition can be lyophilized. These compounds and cells can be adjusted to an appropriate concentration, and optionally combined with other agents. The absolute weight of a given compound and / or other agent included in a unit dose can vary widely. For example, about 0.01 to about 2 g, or about 0.1 to about 500 mg, of at least one compound can be administered. Alternatively, the unit dosage can vary from about 0.01 g to about 50 g, from about 0.01 g to about 35 g, from about 0.1 g to about 25 g, from about 0.5 g to about 12 g, from about 0.5 g to about 8 g, from about 0.5 g to about 4 g, or from about 0.5 g to about 2 g.

[1147] Daily doses of the compounds can vary as well. Such daily doses can range, for example, from about 0.1 g / day to about 50 g / day, from about 0.1 g / day to about 25 g / day, from about 0.1 g / day to about 12 g / day, from about 0.5 g / day to about 8 g / day, from about 0.5 g / day to about 4 g / day, and from about 0.5 g / day to about 2 g / day.

[1148] It will be appreciated that the amount of compounds and cells for use in treatment will vary not only with the particular carrier selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient. Ultimately, the attendant health care provider may determine proper dosage. A pharmaceutical composition may be formulated with the appropriate ratio of each compound in a single unit dosage form for administration with or without cells. Cells can be separately provided and either mixed with a liquid solution of the compound composition, or administered separately. The compounds can also be formulated for sustained release (for example, using microencapsulation, see WO 94 / 07529, and U.S. Pat. No. 4,962,091). The formulations may, where appropriate, be conveniently presented in discrete unit dosage forms and may be prepared by any of the methods well known to the pharmaceutical arts. Such methods may include the step ofmixing the therapeutic agent with liquid carriers, solid matrices, semi-solid carriers, finely divided solid carriers or combinations thereof, and then, if necessary, introducing or shaping the product into the desired delivery system. One or more suitable unit dosage forms containing the compounds and / or the reprogrammed cells can be administered by a variety of routes including parenteral (including subcutaneous, intravenous, intramuscular and intraperitoneal), intracranial, intraspinal, oral, rectal, dermal, transdermal, intrathoracic, intrapulmonary and intranasal (respiratory) routes.

[1149] In one embodiment iPSC are generated from various sources and utilized to generate insulin producing cells in vitro. In other embodiments the process is performed in vivo. Methods for generation of iPSCs from somatic cells involves forced expression of a set of polypeptides or induction factors (IFs). IFs currently known to the art include but are not limited to polypeptides encoded by the genes: c-Myc, Oct3 / 4, Sox2, and Klf4. In addition, small molecule compounds such as histone deacetylace inhibitors may be used or a combination of IFs and small molecules may be used to generate iPSCs. The somatic cells may be used directly, i.e., without culturing or passaging, in the referenced induction methods; or, the somatic cells may be cultured and / or passaged prior to their use in the referenced induction methods. The induced cells may be induced from the somatic cells of a postnatal donor or non-pluripotent donor as described in U.S. application Ser. No. 12 / 157,967, filed Jun. 13, 2008; First Inventor Kazuhiro Sakurada, which is herein incorporated by reference in its entirety. The induced stem cells may be generated from any cell-type including but not limited to those described.

[1150] IPSCs or iSCs may be used directly for differentiation or regenerative medicine. In other cases, iPSCs or iSCs may be stored by the regenerative medicine business, stem cell technology business or a third party. Alternatively, iPSCs or iSCs may be expanded using culturing methods described in U.S. application Ser. No. 12 / 157,967, filed Jun. 13, 2008; First Inventor Kazuhiro Sakurada, which is herein incorporated by reference in its entirety, prior to or after storage. iSCs may be stored in any manner which preserves their multipotent or pluripotent capabilities including cryogenic storage, and culturing. In some cases the donor, potential recipient of the iPSCs or derivatives thereof, or payee may be billed for generation and or delivery of iPSCs or differentiated cells or tissues. In some cases a kit may be marketed and sold which includes a means for generation of iPSCs. During the induction process, forced expression of certain polypeptides is carried out in cultured cells for a period of time, after which the induced cells are screened for anumber of morphological and gene expression properties that characterize multipotent and pluripotent stem cells. Induced cells that meet these screening criteria may then be subcloned and expanded. In some cases, the cells to be induced may be cultured for a period of time prior to the induction procedure. Alternatively, the cells to be induced may be used directly in the induction process without a prior culture period. In some embodiments, the type of cell culture medium used is the same or very similar before, during, and after the induction process. In other cases, different cell culture media are used at different points. For example, one type of culture medium may be used directly before the induction process, while a second type of media is used during the induction process. At times, a third type of culture medium is used during the induction process. Cells may be cultured in medium supplemented with a particular serum. In some embodiments, the serum is fetal bovine serum (FBS). The serum can also be fetal calf serum (FCS). In some cases, the serum may be Human AB serum. Mixtures of serum may also be used, e.g. mixture of FBS and Human AB, FBS and FCS, or FCS and Human AB. Culture of cells may be carried out under a low serum culture conditions prior to, during, or following induction. A "low serum culture condition" refers to the use of a cell culture medium containing a concentration of serum ranging from 0% (v / v) (i.e., serum-free) to about 5% (v / v), e.g., 0% to 2%, 0% to 2.5%, 0% to 3%, 0% to 4%, 0% to 5%, 0.1% to 2%, 0.1% to 5%, 0.1%, 0.5%, 1%, 1.2%, 1.5%, 2%, 2.5%, 3%, 3.5%, or 4%. In some embodiments, the serum concentration is from about 0% to about 2%. In some cases, the serum concentration is about 2%. In some cases, the serum concentration is preferably 2% or less. In other embodiments, cells are cultured under a "high serum condition," i.e., greater than 5% serum to about 20% serum, e.g., 6%, 7%, 8%, 10%, 12%, 15%, or 20%. Culturing under high serum conditions may occur prior to, during, and / or after induction. Some representative media that the cells can be cultured in include: MAPC, FBM, ES, MEF-conditioned ES (MC-ES), and mTeSR™ (available, e.g., from StemCell Technologies, Vancouver, Canada), See Ludwig et al (2006), Nat Biotechnol, 24(2):185-187. In other cases, alternative culture conditions for growth of human ES cells are used, as described in, e.g., Skottman et al (2006), Reproduction, 132(5):691- 698. In some embodiments, the cells are cultured in MAPC, FBM, MC-ES, or mTeSR™ prior to and / or during the introduction of induction factors to the cells; and the cells are cultured in MC-ES or mTeSR™ medium later in the induction process. MAPC (2% FBS) Medium may comprise: 60% Dulbecco's Modified Eagle's Medium-low glucose, 40% MCDB 201, Insulin Transferrin Selenium supplement, (0.01 mg / ml insulin; 0.0055 mg / mltransferrin; 0.005 pg / ml sodium selenite), lx linolenic acid albumin (1 mg / mL albumin; 2 moles linoneic acid / mole albumin), 1 nM dexamethasone, 2% fetal bovine serum, 1 nM dexamethasone, 10-4 M ascorbic acid, and 10 pg / ml gentamycin. FBM (2% FBS) Medium may comprise: MCDB202 modified medium, 2% fetal bovine serum, 5 pg / ml insulin, 50 mg / ml gentamycin, and 50 ng / ml amphotericin-B. ES Medium may comprise: 40% Dulbecco's Modified Eagle's Medium (DMEM) 40% F12 medium, 2 mM L-glutamine, lx non-essential amino acids (Sigma, Inc., St. Louis, Mo.), 20% Knockout Serum Replacement™ (Invitrogen, Inc., Carlsbad, Calif.), and 10 pg / ml gentamycin. MC-ES medium may be prepared as follows. ES medium is conditioned on mitomycin C-treated murine pluripotent fibroblasts (MEFs), harvested, filtered through a 0.45-pM filter, and supplemented with about 0.1 mM mercaptoethanol, about 10 ng / ml bFGF or FGF-2, and, optionally, about 10 ng / ml activin A. In some cases, irradiated MEFs are used in place of the mitomycin C-treated MEFs. When either low or high serum conditions are used for culturing the cells, one or more growth factors such as fibroblast growth factor (FGF)-2; basic FGF (bFGF); platelet-derived growth factor (PDGF), epidermal growth factor (EGF); insulin-like growth factor (IGF); or insulin can be included in the culture medium. Other growth factors that can be used to supplement cell culture media include, but are not limited to one or more: Transforming Growth Factor n-1 (TGF n-1), Activin A, Noggin, Brain-derived Neurotrophic Factor (BDNF), Nerve Growth Factor (NGF), Neurotrophin (NT)-l, NT-2, or NT 3. In some cases, one or more of such factors is used in place of the bFGF or FGF-2 in the MC-ES medium or other cell culture medium. In some cases, the concentration of growth factors in the culture media described (e.g., MAPC, FBM, MC-ES, mTeSR™) is from about 2 ng / ml to about 20 ng / ml, e.g., about 2 ng / ml, 3 ng / ml, 4 ng / ml, 5 ng / ml, 6 ng / ml, 7 ng / ml, 8 ng / ml, 10 ng / ml, 12 ng / ml, 14 ng / ml, 15 ng / ml, 17 ng / ml, or 20 ng / ml. In some embodiments, the concentration of bFGF or FGF2 is from about 2 ng / ml to about 5 ng / ml; from about 5 ng / ml to about 8 ng / ml; from about 9 ng / ml to about 11 ng / ml; from about 11 ng / ml to about 15 ng / ml; or from about 15 ng / ml to about 20 ng / ml. The growth factors may be used alone or in combination. For example, FGF-2 may be added alone to the medium; in another example, both PDGF and EGF are added to the culture medium. In some examples, following initiation of the forced expression of genes or polypeptides (e.g., immediately after a retroviral infection period) in cells, the "induced cells" are maintained in MC-ES medium as described herein.

[1151] In some embodiments, cells are maintained in the presence of a rho, or rho- associated, protein kinase (ROCK) inhibitor to reduce apoptosis. In some cases, an inhibitor of Rho associated kinase is added to the culture medium. For example, the addition of Y-27632 (Calbiochem; water soluble) or Fasudil (HA1077: Calbiochem), an inhibitor of Rho associated kinase (Rho associated coiled coil-containing protein kinase) may be used to culture the human pluripotent and multipotent stem cells of the present invention. In some cases the concentration of Y-27632 or Fasudil, is from about 5 pM to about 20 pM, e.g., about 5 pM, 10 pM, 15 pM, or 20 pM. The cells may be cultured for about 1 to about 12 days e.g., 2 days, 3 days, 4.5 days, 5 days, 6.5 days, 7 days, 8 days, 9 days, 10 days, or any other number of days from about 1 day to about 12 days prior to undergoing the induction methods described herein. In some cases, the induced cells are cultured in complete ES medium in a 37nC, 5% CO2 incubator, with medium changes about every 1 to 2 days. In some embodiments, induced the induced cells are cultured and observed for about 14 days to about 40 days, e.g., 15, 16, 17, 18, 19, 20, 23, 24, 27 , 28, 29, 30, 31, 33, 34, 35, 36, 37, 38 days, or any other period from about 14 days to about 40 days prior to identifying and selecting clones comprising "induced cells" based on morphological characteristics. Morphological characteristics for identifying induced cell clones include, but are not limited to, a small cell size with a high nucleus-to- cytoplasm ratio; formation of small monolayer colonies within the space between parental cells (e.g., between fibroblasts). The cells may be plated at a cell density of about 1x103 cells / cm2 to about 1x104 cells / cm2, e.g., 2x103 cells / cm2, 3.5x103 cells / cm2, 6x103 cells / cm2, 7x103 cells / cm2, 9x103 cells / cm2, or any other cell density from about 1x103 cells / cm2 to about 1x104 cells / cm2. The cells can be plated and cultured directly on tissue culture-grade plastic. Alternatively, cells are plated and cultured on a coated substrate, e.g., a substrate coated with fibronectin, gelatin, matrigel™, collagen, or laminin. Suitable cell culture vessels include, e.g., 35 mm, 60 mm, 100 mm, and 150 mm cell culture dishes, 6-well cell culture plates, and other sizeequivalent cell culture vessels. In some cases, the cells are cultured with feeder cells. For example, the cells may be cultured on a layer, or carpet, of MEFs.

[1152] Media with low concentrations of serum may be particularly useful to enrich for undifferentiated stem cells. The undifferentiated cells cultured under low serum conditions may or may not share certain properties with MSCs, MAPCs, and / or MIAMI cells. Differences in phenotype may be due, in part, to culture methods used to obtain MSCs, MAPCs and MIAMI cells. For example, MSCs are often obtained by isolating thenon-hematopoeitic cells (e.g., interstitial cells) adhering to a plastic culture dish when tissue, e.g., bone marrow, fat, muscle, or skin etc., is cultured in a culture medium containing a high-concentration serum (5% or more). However, even under these culture conditions, a very small number of undifferentiated cells can be maintained, especially if the cells were passaged under certain culture conditions (e.g., low passage number or low-density culturing). In some embodiments, in order to culture and grow human pluripotent stem cells induced from the undifferentiated stem cells of the present invention present in a human postnatal tissue, it is preferred that the cells are subcultured every 5 to 7 days in a culture medium containing the additives described herein on a MEF-covered plastic culture dish or a matrigel-coated plastic culture dish. In some cases, the cells may be cultured at a low density, which may be accomplished by splitting the cells from about 1:6 to 1:3 or by plating the cells at 103 cells / cm2 to 3x104 cells / cm2. Primary culture ordinarily occurs immediately after the cells are isolated from a donor, e.g., human. The primary cells can be subjected to a second subculture, a third subculture, a fourth subculture, and greater than four subcultures. A "second" subculture describes primary culture cells subcultured once, a "third" subculture describes primary cultures subcultured twice, a "fourth" subculture describes primary cells subcultured three times, etc. The culture techniques described herein may generally include culturing from the period between the primary culture and the fourth subculture, but other culture periods may also be employed. Preferably, cells are cultured from primary culture to second subculture.

[1153] Inducing a cell to become multipotent or pluripotent can be accomplished in numerous ways. In some embodiments, the methods for induction of pluripotency or multipotency in one or more cells include forcing expression of a set of induction factors (IFs). In some cases, the set of IFs includes one or more: an Oct3 / 4 polypeptide, a Sox2 polypeptide, a Klf4 polypeptide, or a c-Myc polypeptide. In some cases, the set does not include a c-Myc polypeptide. For example, the set of IFs can include: an Oct3 / 4 polypeptide, a Sox2 polypeptide, and a Klf4 polypeptide, but not a c-Myc polypeptide. In some cases, the set of IFs does not include polypeptides that might increase the risk of cell transformation. In some cases, the set may include a c-Myc polypeptide. In certain cases, the c-Myc polypeptide is a constitutively active variant of c-Myc. In some instances, the set includes a c-Myc polypeptide capable of inducible activity, e.g., a c- Myc-ER polypeptide,

[1154] In other cases, the set of IFs may include: an Oct3 / 4 polypeptide, a Sox2 polypeptide, and a Klf4 polypeptide, but not a TERT polypeptide, a SV40 Large T antigen polypeptide, HPV16 E6 polypeptide, a HPV16 E7 polypeptide, or a Bmil polypeptide. In some cases, the set of IFs does not include a TERT polypeptide. In some cases, the set of IFs does not include a SV40 Large T antigen. In other cases, the set of IFS does not include a HPV 16 E6 polypeptide or a HPV 16 E7 polypeptide.

[1155] In some cases, the set of IFs includes three IFs, wherein two of the three IFs are an Oct3 / 4 polypeptide and a Sox2 polypeptide. In other cases, the set of IFs includes two IFs, wherein the two polypeptides are a c-Myc polypeptide and a Sox2 polypeptide In some cases, the set of induction factors is limited to Oct 3 / 4, Sox2, and Klf4 polypeptides. In other cases, the set of induction factors may be limited to a set of four IFs: an Oct3 / 4 polypeptide, a Sox2 polypeptide, a Klf4 polypeptide, and a c-Myc polypeptide. A set of IFs may include IFs in addition to an Oct 3 / 4, a Sox2, and a Klf4 polypeptide. Such additional IFs include, but are not limited to Nanog, TERT, LIN28, CYP26A1, GDF3, FoxD3, Zfp42, Dnmt3b, Ecatl, and Tell polypeptides. In some cases, the set of additional IFs does not include a c Myc polypeptide. In some cases, the set of additional IFs does not include polypeptides that might increase the risk of cell transformation. Forced expression of IFs may be maintained for a period of at least about 7 days to at least about 40 days, e.g., 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 25 days, 30 days, 33 days, or 37 days. In a first step, a cell population (the starting cell population) comprising at least one cell capable of differentiation is provided. In some embodiments, the cell capable of differentiation is a pancreatic progenitor cell expressing PDX1 and NKX6.1.

[1156] In some embodiments, the starting cell population comprises at least 5% pancreatic progenitor cells, such as at least 10% pancreatic progenitor cells, such as at least 15% pancreatic progenitor cells, such as at least 20% pancreatic progenitor cells, such as at least 25% pancreatic progenitor cells, such as at least 30% pancreatic progenitor cells, such as at least 35% pancreatic progenitor cells, such as at least 40% pancreatic progenitor cells, such as at least 45% pancreatic progenitor cells, such as at least 50% pancreatic progenitor cells, such as at least 55% pancreatic progenitor cells, such as at least 60% pancreatic progenitor cells, such as at least 65 pancreatic progenitor cells, such as at least 70% pancreatic progenitor cells, such as at least 75% pancreatic progenitor cells, such as at least 80% pancreatic progenitor cells, such as at least 85%pancreatic progenitor cells, such as at least 90% pancreatic progenitor cells, such as at least 95% pancreatic progenitor cells. In order to determine the fraction of progenitor cells comprised in a cell population, for example in the starting population, methods known in the art can be employed, such as, but not limited to, immunostaining or flow cytometry methods.

[1157] Without being bound by theory, the percentage of pancreatic progenitor cells in the starting cell population can be estimated by the expression of GP2. Thus in some embodiments, the starting cell population comprises at least 5% cells expressing GP2, such as at least 10% cells expressing GP2, such as at least 15% cells expressing GP2, such as at least 20% cells expressing GP2, such as at least 25% cells expressing GP2, such as at least 30% cells expressing GP2, such as at least 35% cells expressing GP2, such as at least 40% cells expressing GP2, such as at least 45% cells expressing GP2, such as at least 50% cells expressing GP2, such as at least 55% cells expressing GP2, such as at least 60% cells expressing GP2, such as at least 65% cells expressing GP2, such as at least 70% cells expressing GP2, such as at least 75% cells expressing GP2, such as at least 80% cells expressing GP2, such as at least 85% cells expressing GP2, such as at least 90% pancreatic progenitor cells, such as at least 95% pancreatic progenitor cells. GP2 expression can be determined by methods known in the art, such as immunostaining methods, flow cytometry methods or quantitative measurements of transcription levels. Likewise, without being bound by theory, the percentage of PDX1+NKX6.1+ cells in the starting cell population can be estimated by the expression of GP2. Thus in some embodiments, the starting cell population comprises at least 5% cells expressing GP2, such as at least 10% cells expressing GP2, such as at least 15% cells expressing GP2, such as at least 20% cells expressing GP2, such as at least 25% cells expressing GP2, such as at least 30% cells expressing GP2, such as at least 35% cells expressing GP2, such as at least 40% cells expressing GP2, such as at least 45% cells expressing GP2, such as at least 50% cells expressing GP2, such as at least 55% cells expressing GP2, such as at least 60% cells expressing GP2, such as at least 65% cells expressing GP2, such as at least 70% cells expressing GP2, such as at least 75% cells expressing GP2, such as at least 80% cells expressing GP2, such as at least 85% cells expressing GP2, such as at least 90% pancreatic progenitor cells, such as at least 95% pancreatic progenitor cells. GP2 expression can be determined by methods known in the art, such as immunostaining methods, flow cytometry methods or quantitative measurements of transcription levels.

[1158] In some embodiments, the cell population may be derived or isolated from an individual, such as, but not limited to, a mammal, for example a human. In some embodiments, the cells capable of differentiation are pluripotent stem cells, for example human pluripotent stem cells (hPSCs). hPSCs include human induced pluripotent stem cells (hiPSCs and naive human stem cells (NhSCs). In one embodiment, the starting cell population is obtained from a pancreas, including a foetal pancreas or an adult pancreas. In one aspect, the pancreas is from a mammal, such as a human. In another embodiment, the starting cell population is a somatic cell population. In some embodiments, the starting cell population comprises at least one pancreatic progenitor cell expressing PDX1 and NKX6.1 and is obtained from a somatic cell population. In a further aspect of the invention, the somatic cell population has been induced to dedifferentiate into an pluripotent-like stem cell (ESC, e.g. a pluripotent stem cell, or hESCs for human ESCs). Such dedifferentiated cells are also termed induced pluripotent stem cells (IPSCs, or hiPSCs for human IPSCs). In yet another embodiment, the starting cell population is ESCs or hESCs. In one embodiment, the starting cell population is obtained from ESCs or hESCs. In some embodiments, the starting cell population is a population of pluripotent stem cells such as ESC like-cells. In some embodiments, a cell population comprising at least one pancreatic progenitor cell may be obtained by methods known in the art, before steps viii) and ix) as described herein are performed. For example, differentiation can be induced in embryoid bodies and / or in monolayer cell cultures or a combination thereof. In one aspect of the invention, the starting cell population is of mammalian origin. In one aspect of the invention, the starting cell population is of human origin.Example 1: Generation of Cardiogenic Progenitors from Inducible Pluripotent Stem Cells (Alpha Myosin Heavy Chain)

[1159] Fibroblast derived induced pluripotent stem cells were purchased from ATCC and cultured according to the manufacturer's instructions. Cells were plated in either purchased cardiogenic media (Stem Cell Technologies Inc, Vancouver Canada) or conditioned media generated from decellularized cardiac tissue seeded with mesenchymal stem cells in the presence of 100 pg / ml valproic acid. Cells are cultured forthe time period indicated below. Expression of alpha myosin heavy chain was assessed by Western Blot and expressed as percentage of control. Results are shown in FIG. 1.Example 2: Generation of Cardiogenic Progenitors from Inducible Pluripotent Stem Cells (c- kit)

[1160] Fibroblast derived induced pluripotent stem cells were purchased from ATCC and cultured according to the manufacturer's instructions. Cells were plated in either purchased cardiogenic media (Stem Cell Technologies Inc, Vancouver Canada) or conditioned media generated from decellularized cardiac tissue seeded with mesenchymal stem cells. Expression of c-kit on troponin expressing cells was assessed by Western Blot and expressed as percentage of control. Results are shown in FIG. 2.Example 3: Generation of Cardiogenic Progenitors from Inducible Pluripotent Stem Cells (CD133)

[1161] Fibroblast derived induced pluripotent stem cells were purchased from ATCC and cultured according to the manufacturer's instructions. Cells were plated in either purchased cardiogenic media (Stem Cell Technologies Inc, Vancouver Canada) or conditioned media generated from decellularized cardiac tissue seeded with mesenchymal stem cells. Expression of CD133 on troponin expressing cells was assessed by Western Blot and expressed as percentage of control. Results are shown in FIG. 3.

Claims

Claims1. A method of generating tissue specific progenitors from pluripotent stem cells comprising exposing said pluripotent stem cells to factors generated by representative tissue cells.

2. The method of claim 1, wherein said pluripotent stem cells are capable of forming a teratoma in an immune deficient mouse.

3. The method of claim 1, wherein said pluripotent stem cells are generated by induced pluripotency.

4. The method of claim 3, wherein said pluripotency is induced by administration of a pluripotency factor together with one or more dedifferentiation factors.

5. The method of claim 4, wherein said dedifferentiation factor is conditioned media from an embryonic stem cell.

6. The method of claim 5, wherein said dedifferentiation factor is conditioned media from a pluripotent stem cell that has been exposed to cellular stress.

7. The method of claim 6, wherein said cellular stress is selected from a group of stressors consisting of: a) hypoxia; b) metabolic stress; c) hyperthermia; d) hypothermia; e) hypotonic stress; f) hypertonic stress; and e) proteosome blockade stress.

8. The method of claim 1, wherein said factors generated by representative tissue cells are factors derived from decellularized tissue.

9. The method of claim 8, wherein said decellularized tissue is administered to culture media containing pluripotent stem cells.

10. A method of generating cardiogenic progenitor cells comprising of culturing a pluripotent stem cell with a media containing decellularized cardiac tissue.

11. The method of claim 10, wherein said pluripotent stem cells are induced pluripotent stem cells.

12. The method of claim 10, wherein said media contains valproic acid.

13. The method of claim 12, wherein said valproic acid is added to said media at a concentration of 10 pg / ml to 100 ng / ml.

14. The method of claim 13, wherein said valproic acid is added to said media at a concentration of 100 pg / ml to 10 ng / ml.

15. A method of generating cardiogenic progenitor cells comprising of culturing a pluripotent stem cell with a cardiogenic media combined with mesenchymal stem cell conditioned media.

16. The method of claim 15, wherein said cardiogenic media is mixed with stem cell conditioned media generated by growing stem cells on decellularized bone matrix.

17. The method of claim 16, wherein said stem cell conditioned media is generated by culture of mesenchymal stem cells derived from bone marrow on bone decellularized matrix.

18. The method of claim 15, wherein said cardiogenic media contains valporoic acid.

19. The method of claim 15, wherein said valproic acid is added to said media at a concentration of 10 pg / ml to 100 ng / ml.

20. The method of claim 15, wherein said valproic acid is added to said media at a concentration of 100 pg / ml to 10 ng / ml.

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