Generation of personalized mesenchymal stem cells

Abstract

Novel methods of creating personalized mesenchymal stem cells from pluripotent stem cells, including the utilization of extracellular matrix components, optionally seeded with exogenous differentiation factors for creation of a substrate for promotion of mesenchymal stem cell formation from pluripotent cell populations. Matrices useful for stimulation of differentiation include decellularized bone matrix, placental matrix, and adipose tissue derived matrix. Growth factors such as BMP2, BMP4, GDF-11, and stem cell factor can be added to said matrix. Stimulation of differentiation can be induced by exposure to conditioned media isolated from mesenchymal stem cells.

Description

GENERATION OF PERSONALIZED MESENCHYMAL STEM CELLSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims benefit of U.S. Provisional Patent Application Serial No. 63 / 599,431, filed on November 15, 2023, entitled "Generation of Personalized Mesenchymal Stem Cells", the contents of which are incorporated herein by reference in its entirety.FIELD OF THE INVENTION

[0002] The invention belongs to the field of regenerative medicine, more specifically, the invention belongs to the field of generation of functional therapeutic cells from pluripotent stem cells, more specifically, the invention belongs to the field of generating mesenchymal stem cells from pluripotent stem cells.BACKGROUN D OF TH E INVENTION

[0003] Several therapeutic cell types are known in the art including hematopoietic stem cells, immunological cells, and mesenchymal stem cells (MSC). MSC were originally found in bone marrow and possess high potential in regenerative medicine based on ability to exert therapeutic activities in a variety of allogeneic and autologous situations. MSCs may be differentiated into various types of in vivo mesoderm lineages, for example, osteocytes, chondrocytes, tendinocytes, adipocytes, myocytes, fibroblasts, and the like. Also, MSCs may be trans-differentiated into nerve cells, myocardial cells, endothelial cells, and interstitial cells under appropriate medium conditions. In addition, bone marrow MSCs express class I MHC antigens other than class II MHC antigens or express co-stimulatory molecules indicating that the MSCs have no immunogenic activities (Klyushnenkova E. et al., J Biomed Sci, 12(1): 47-57, 2005). In addition, because MSCs exhibit immune modulatory activities, the cells may be used as graft enhancers or inhibitors for fatal graft and host diseases (Le Blanc K et al., Lancet, 363 (9419): 1439- 1441, 2004; El-Badri N. S et al., Exp Hematol, 26(2): 110-116, 1998). Such MSCs may be isolated from various adult tissues such as bone marrow, adipose tissues, cord blood, peripheral blood, neonatal tissues, human placenta, and the like, but have a limitation in the number of MSCs obtained from the adult tissues.

[0004] The current invention provides means of generating MSCs from pluripotent personalized sources, the immortal nature of the originator cell population allows for numerous modifications to be made including gene modification, gene editing and gene silencing.SUMMARY OF THE INVENTION

[0005] A summary is provided below based on numbered aspects of the invention.

[0006] 1. A method of generating mesenchymal stem cells comprising the steps of: a) obtaining a pluripotent stem cell; b) contacting said pluripotent stem cell with a matrix inductive of mesenchymal stem cell differentiation; c) optionally seeding said matrix with mesenchymal stem cell stimulatory growth factors; and d) isolating said resulting mesenchymal stem cells.

[0007] 2. The method of aspect 1, wherein said pluripotent stem cell expresses hTERT.

[0008] 3. The method of aspect 1, wherein said pluripotent stem cell is an induced pluripotent stem cell.

[0009] 4. The method of aspect 1, wherein said matrix inductive of mesenchymal stem cell differentiation is decellularized bone.

[0010] 5. The method of aspect 4, wherein said decellularized bone possesses intactTGF-beta molecules.

[0011] 6. The method of aspect 5, wherein said decellularized bone contains FGF-1 at10-1000 pg of FGF-1 per gram of bone tissue.

[0012] 7. The method of aspect 5, wherein said decellularized bone contains FGF-1 at50-500 pg of FGF-1 per gram of bone tissue.

[0013] 8. The method of aspect 5, wherein said decellularized bone contains FGF-1 at100-250 pg of FGF-1 per gram of bone tissue.

[0014] 9. The method of aspect 5, wherein said decellularized bone contains BMP-2 at100-5000 pg of BMP-2 per gram of bone tissue.

[0015] 10. The method of aspect 5, wherein said decellularized bone contains BMP-2 at500-2500 pg of BMP-2 per gram of bone tissue.

[0016] 11. The method of aspect 5, wherein said decellularized bone contains BMP-2 at1000-2000 pg of BMP-2 per gram of bone tissue.

[0017] 12. The method of aspect 4, wherein said decellularized bone possesses intactMMP3 molecules.

[0018] 13. The method of aspect 4, wherein said decellularized bone possesses intactMMP5 molecules.

[0019] 14. The method of aspect 4, wherein said decellularized bone possesses intactMMP7 molecules.

[0020] 15. The method of aspect 4, wherein said decellularized bone possesses intactMMP9 molecules.

[0021] 16. The method of aspect 4, wherein said decellularized bone possesses intact tissue inhibitor of matrix metalloprotease molecules.

[0022] 17. The method of aspect 4, wherein said decellularized bone possesses intact granulocyte colony stimulating factor.

[0023] 18. The method of aspect 4, wherein said decellularized bone possesses intact granulocyte monocyte colony stimulating factor.

[0024] 19. The method of aspect 4, wherein said decellularized bone possesses intact macrophage colony stimulating factor.

[0025] 20. The method of aspect 4, wherein said decellularized bone possesses intact angiopoietin.

[0026] 21. The method of aspect 1, wherein said pluripotent stem cells are allowed to form embryoid bodies before contacting with said mesenchymal stem cell inductive matrix.

[0027] 22. The method of aspect 21, wherein said embryoid bodies are formed by culture of said pluripotent stem cells in absence of feeder cells.

[0028] 23. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of leukemia inhibitory factor.

[0029] 24. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of interleukin-3.

[0030] 25. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of interleukin-6.

[0031] 26. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of interleukin-11.

[0032] 27. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of interleukin-35.

[0033] 28. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of interleukin-37.

[0034] 29. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of steel factor.

[0035] 30. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of osteopontin.

[0036] 31. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of thrombopoietin.

[0037] 32. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of hepatocyte growth factor.

[0038] 33. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of transforming growth factor beta.

[0039] 34. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of endoglin.

[0040] 35. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of epidermal growth factor.

[0041] 36. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of amphiregulin.

[0042] 37. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of VEGF.

[0043]

[0044] 38. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of VEGF-C.

[0045] 39. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of fibroblast conditioned media.

[0046] 40. The method of aspect 22, wherein said culture in absence of feeder cells is performed in the presence of mesenchymal stem cell conditioned media.

[0047] 41. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 40 ng / ml of VEGF.

[0048] 42. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 100 pg / ml of FGF-1.

[0049] 43. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of FGF-2.

[0050] 44. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 1 ng / ml of FGF-5.

[0051] 45. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 100 pg / ml of amphiregulin.

[0052] 46. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 100 pg / ml of IIGF-1.

[0053] 47. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of IGF-BP.

[0054] 48. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 10 pg / ml of endoglin.

[0055] 49. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 100 pg / ml of CXCL12.

[0056] 50. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 10 pg / ml of MIP-l beta.

[0057] 51. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 80 ng / ml of VEGF.

[0058] 52. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of FGF-1.

[0059] 53. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 400 pg / ml of FGF-2.

[0060] 54. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 2 ng / ml of FGF-5.

[0061] 55. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of amphiregulin.

[0062] 56. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of IIGF-1.

[0063] 57. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 400 pg / ml of IGF-BP.

[0064] 58. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 20 pg / ml of endoglin.

[0065] 59. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 200 pg / ml of CXCL12.

[0066] 60. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 20 pg / ml of MIP-l beta.

[0067] 61. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 160 ng / ml of VEGF.

[0068] 62. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 400 pg / ml of FGF-1.

[0069] 63. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 800 pg / ml of FGF-2.

[0070] 64. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 4 ng / ml of FGF-5.

[0071] 65. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 400 pg / ml of amphiregulin.

[0072] 66. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 400 pg / ml of IIGF-1.

[0073] 67. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 800 pg / ml of IGF-BP.

[0074] 68. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 40 pg / ml of endoglin.

[0075] 69. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 700 pg / ml of CXCL12.

[0076] 70. The method of aspect 40, wherein said mesenchymal stem cell conditioned media contains at least 40 pg / ml of MIP-l beta.

[0077] 71. The method of aspect 40, wherein said mesenchymal stem cell conditioned media is generated by treatment of mesenchymal stem cells with a cellular stressor.

[0078] 72. The method of aspect 71, wherein said cellular stressor is hypoxia.

[0079] 73. The method of aspect 71, wherein said cellular stressor is hyperoxia.

[0080] 74. The method of aspect 71, wherein said cellular stressor is hyperthermia.

[0081] 75. The method of aspect 71, wherein said cellular stressor is exposure to hypertonic conditions.

[0082] 76. The method of aspect 71, wherein said cellular stressor is exposure to hypotonic conditions.

[0083] 77. The method of aspect 71, wherein said cellular stressor is exposure to xenon.

[0084] 78. The method of aspect 71, wherein said cellular stressor is exposure to hydrogen sulfide.

[0085] 79. The method of aspect 71, wherein said cellular stressor is exposure to hydrogen gas.

[0086] 80. The method of aspect 71, wherein said cellular stressor is exposure to an inducer of heme-oxygenase-1.

[0087] 81. The method of aspect 80, wherein said inducer of heme-oxygenase-1 is carbon monoxide.

[0088] 82. The method of aspect 71, wherein said cellular stressor is exposure to an activator of NF-kappa B.

[0089] 83. The method of aspect 82, wherein said activator of NF-kappa B is HMGBl.

[0090] 84. The method of aspect 82, wherein said activator of NF-kappa B is interleukin-1 beta.

[0091] 85. The method of aspect 82, wherein said activator of NF-kappa B is an agonist of toll like receptor 2.

[0092] 86. The method of aspect 85, wherein said agonist of said toll like receptor 2 is peptidoglycan.

[0093] 87. The method of aspect 82, wherein said activator of NF-kappa B is zymosan.

[0094] 88. The method of aspect 82, wherein said activator of NF-kappa B is beta glucan.

[0095] 89. The method of aspect 82, wherein said activator of NF-kappa B is BCG.

[0096] 90. The method of aspect 82, wherein said activator of NF-kappa B is Reptimed.

[0097] 91. The method of aspect 82, wherein said activator of NF-kappa B is TNF-alpha.

[0098] 92. The method of aspect 82, wherein said activator of NF-kappa B is LIGHT.

[0099] 93. The method of aspect 82, wherein said activator of NF-kappa B is BLyS.

[0100] 94. The method of aspect 82, wherein said activator of NF-kappa B is TRANCE.

[0101] 95. The method of aspect 82, wherein said activator of NF-kappa B is RANK ligand.

[0102] 96. The method of aspect 82, wherein said activator of NF-kappa B is Fas ligand.

[0103] 97. The method of aspect 82, wherein said activator of NF-kappa B is interferon gamma.

[0104] 98. The method of aspect 82, wherein said activator of NF-kappa B is Poly IC.

[0105] 99. The method of aspect 82, wherein said activator of NF-kappa B is Poly LC.

[0106] 100. The method of aspect 82, wherein said activator of NF-kappa B is low molecular weight hyaluronic acid.

[0107] 101. The method of aspect 82, wherein said activator of NF-kappa B is double stranded RNA.

[0108] 102. The method of aspect 82, wherein said activator of NF-kappa B is bacterialDNA.

[0109] 103. The method of aspect 82, wherein said activator of NF-kappa B is DNA containing CpG motifs.

[0110] 104. The method of aspect 82, wherein said activator of NF-kappa B is circularDNA.

[0111] 105. The method of aspect 82, wherein said activator of NF-kappa B is uric acid crystals.

[0112] 106. The method of aspect 1, wherein said mesenchymal stem cell is plastic adherent.

[0113] 107. The method of aspect 1, wherein said mesenchymal stem cell possesses a fibroblastoid-like morphology.

[0114] 108. The method of aspect 1, wherein said mesenchymal stem cell is capable of undergoing orthodox differentiation.

[0115] 109. The method of aspect 108, wherein said orthodox differentiation is conversion into chondrocytes.

[0116] 110. The method of aspect 109, wherein said chondrocytes express type 2 collagen.

[0117] 111. The method of aspect 110, wherein said chondrocytes are generated as a result of hydrostatic pressure on said mesenchymal stem cells.

[0118] 112. The method of aspect 110, wherein said chondrocytes are generated as a result of BMP2 treatment of said mesenchymal stem cells.

[0119] 113. The method of aspect 110, wherein said chondrocytes are generated as a result of BMP2 and TGF-beta treatment of said mesenchymal stem cells.

[0120] 114. The method of aspect 110, wherein said chondrocytes are generated as a result of TGF-beta treatment of said mesenchymal stem cells.

[0121] 115. The method of aspect 110, wherein said chondrocytes are generated as a result of FGF-2 and BMP-2 treatment of said mesenchymal stem cells.

[0122] 116. The method of aspect 110, wherein said chondrocytes are generated as a result of BMP4 treatment of said mesenchymal stem cells.

[0123] 117. The method of aspect 110, wherein said chondrocytes are generated as a result of SOX9 transfection of said mesenchymal stem cells.

[0124] 118. The method of aspect 110, wherein said chondrocytes are generated as a result of PIM1 transfection of said mesenchymal stem cells.

[0125] 119. The method of aspect 110, wherein said chondrocytes are generated as a result of KLF4 transfection of said mesenchymal stem cells.

[0126] 120. The method of aspect 110, wherein said chondrocytes are generated as a result of IGF-1 transfection of said mesenchymal stem cells.

[0127] 121. The method of aspect 109, wherein said chondrocytes express aggrecan.

[0128] 122. The method of aspect 109, wherein said chondrocytes express glycosaminoglycans.

[0129] 123. The method of aspect 109, wherein said chondrocytes express decay accelerating factor.

[0130] 124. The method of aspect 109, wherein said chondrocytes express elastin.

[0131] 125. The method of aspect 109, wherein said chondrocytes express vimentin.

[0132] 126. The method of aspect 109, wherein said chondrocytes express calreticulin.

[0133] 127. The method of aspect 109, wherein said chondrocytes express complement factor H.

[0134] 128. The method of aspect 109, wherein said chondrocytes express high molecular weight hyaluronic acid.

[0135] 129. The method of aspect 109, wherein said chondrocytes express low levels ofHLA I as compared to fibroblasts.

[0136] 130. The method of aspect 109, wherein said chondrocytes resemble nucleus pulposus cells.

[0137] 131. The method of aspect 108, wherein said orthodox differentiation is conversion into osteocytes.

[0138] 132. The method of aspect 108, wherein said orthodox differentiation is conversion into adipocytes.

[0139] 133. The method of aspect 108, wherein said orthodox differentiation is conversion into myocytes.

[0140] 134. The method of aspect 1, wherein said mesenchymal stem cell is capable of undergoing non-orthodox differentiation.

[0141] 135. The method of aspect 134, wherein non-orthodox differentiation is conversion into insulin producing cells.

[0142] 136. The method of aspect 135, wherein said insulin producing cells expressISLA-1.

[0143] 137. The method of aspect 135, wherein said insulin producing cells expressPDX-1.

[0144] 138. The method of aspect 135, wherein said insulin producing cells express pro-insulin.

[0145] 139. The method of aspect 135, wherein said insulin producing cells express C- peptide.

[0146] 140. The method of aspect 135, wherein said insulin producing cells expressGAD65.

[0147] 141. The method of aspect 135, wherein said insulin producing cells expressGAD67.

[0148] 142. The method of aspect 134, wherein non-orthodox differentiation is conversion into dopamine producing cells.

[0149] 143. The method of aspect 134, wherein non-orthodox differentiation is conversion into neuronal cells.

[0150] 144. The method of aspect 143, wherein said neuronal cells possess NMDA receptor.

[0151] 145. The method of aspect 143, wherein said neuronal cells express nestin.

[0152] 146. The method of aspect 143, wherein said neuronal cells express NeuroD.

[0153] 147. The method of aspect 143, wherein said neuronal cells express GFAP.

[0154] 148. The method of aspect 143, wherein said neuronal cells express dopamine receptor.

[0155] 149. The method of aspect 143, wherein said neuronal cells express serotonin receptor.

[0156] 150. The method of aspect 143, wherein said neuronal cells express glutamine receptor.

[0157] 151. The method of aspect 143, wherein said neuronal cells express GABA receptor.

[0158] 152. The method of aspect 134, wherein non-orthodox differentiation is conversion into hepatocytes.

[0159] 153. The method of aspect 152, wherein said hepatocytes possess detoxification properties.

[0160] 154. The method of aspect 152, wherein said hepatocytes produce albumin.

[0161] 155. The method of aspect 134, wherein non-orthodox differentiation is conversion into type 1 pulmonary epithelial cells.

[0162] 156. The method of aspect 134, wherein non-orthodox differentiation is conversion into type II pulmonary epithelial cells.

[0163] 157. The method of aspect 134, wherein non-orthodox differentiation is conversion into renal epithelial cells.

[0164] 158. The method of aspect 134, wherein non-orthodox differentiation is conversion into pigmented retinal epithelial cells.

[0165] 159. The method of aspect 134, wherein non-orthodox differentiation is conversion into angioblasts.

[0166] 160. The method of aspect 134, wherein non-orthodox differentiation is conversion into endothelial progenitor cells.

[0167] 161. The method of aspect 160, wherein said endothelial progenitor cells are capable of reducing damage to a blood vessel.

[0168] 162. The method of aspect 161, wherein said damage to said blood vessel is restenosis.

[0169] 163. The method of aspect 161, wherein said damage to said blood vessel is neointimal proliferation.

[0170] 164. The method of aspect 161, wherein said damage to said blood vessel is atherosclerosis.

[0171] 165. The method of aspect 161, wherein said damage to said blood vessel is diabetic vision loss.

[0172] 166. The method of aspect 161, wherein said damage to said blood vessel is macular degeneration.

[0173] 167. The method of aspect 161, wherein said damage to said blood vessel is immunologically mediated blood vessel injury.

[0174] 168. The method of aspect 161, wherein said damage to said blood vessel is viral mediated blood vessel injury.

[0175] 169. The method of aspect 161, wherein said damage to said blood vessel isCOVID-19 spike protein mediated blood vessel injury.

[0176] 170. The method of aspect 161, wherein said damage to said blood vessel is vaccine mediated blood vessel injury.

[0177] 171. The method of aspect 161, wherein said damage to said blood vessel isCOVID-19 vaccine mediated blood vessel injury.

[0178] 172. The method of aspect 160, wherein said endothelial progenitor cells express LDL receptor.

[0179] 173. The method of aspect 160, wherein said endothelial progenitor cells express IL-10 receptor.

[0180] 174. The method of aspect 160, wherein said endothelial progenitor cells express interferon gamma receptor.

[0181] 175. The method of aspect 160, wherein said endothelial progenitor cells express IL-6 receptor.

[0182] 176. The method of aspect 160, wherein said endothelial progenitor cells express c-kit.

[0183] 177. The method of aspect 160, wherein said endothelial progenitor cells express CD33.

[0184] 178. The method of aspect 160, wherein said endothelial progenitor cells express CD34.

[0185] 179. The method of aspect 160, wherein said endothelial progenitor cells express CD133.

[0186] 180. The method of aspect 160, wherein said endothelial progenitor cells express CD105.

[0187] 181. The method of aspect 134, wherein non-orthodox differentiation is conversion into hematopoietic stem cells.

[0188] 182. The method of aspect 181, wherein said hematopoietic stem cells expressFas ligand.

[0189] 183. The method of aspect 181, wherein said hematopoietic stem cells expressCD34.

[0190] 184. The method of aspect 181, wherein said hematopoietic stem cells expressNOTCH.

[0191] 185. The method of aspect 181, wherein said hematopoietic stem cells expressWNT-5a.

[0192] 186. The method of aspect 181, wherein said hematopoietic stem cells expressIL-3 receptor.

[0193] 187. The method of aspect 181, wherein said hematopoietic stem cells expressTGF-beta receptor.

[0194] 188. The method of aspect 181, wherein said hematopoietic stem cells expressIL-6 receptor.

[0195] 189. The method of aspect 181, wherein said hematopoietic stem cells expressIL-11 receptor.

[0196] 190. The method of aspect 181, wherein said hematopoietic stem cells expressIL-13 receptor.

[0197] 191. The method of aspect 181, wherein said hematopoietic stem cells expressIL-17 receptor.

[0198] 192. The method of aspect 181, wherein said hematopoietic stem cells expressIL-22 receptor.

[0199] 193. The method of aspect 181, wherein said hematopoietic stem cells expressIL-35 receptor.

[0200] 194. The method of aspect 181, wherein said hematopoietic stem cells expressIL-37 receptor.

[0201] 195. The method of aspect 181, wherein said hematopoietic stem cells express leukemia inhibitory factor receptor.

[0202] 197. The method of aspect 181, wherein said hematopoietic stem cells produceTGF-beta upon stimulation with IL-3.

[0203] 198. The method of aspect 181, wherein said hematopoietic stem cells produceTGF-beta upon stimulation with IL-11.

[0204] 199. The method of aspect 181, wherein said hematopoietic stem cells are capable of inducing multi-lineage hematopoietic engraftment.

[0205] 199. A composition of mesenchymal stem cells generated from a cell reprogrammed to pluripotency.

[0206] 200. The composition of aspect 199, wherein said reprogramming is induced through inhibition of p53 expression.

[0207] 201. The composition of aspect 200, wherein p53 expression is suppression of p53 activity.

[0208] 202. The composition of aspect 201, wherein said suppression of p53 activity is mediated through administration of a small molecule inhibitor of p53.

[0209] 203. The composition of aspect 201, wherein said suppression of p53 activity is mediated through administration of decoy oligonucleotides.

[0210] 204. The composition of aspect 201, wherein said suppression of p53 activity is mediated through administration of decoy peptides.

[0211] 205. The composition of aspect 201, wherein said suppression of p53 expression is achieved through induction of RNA interference targeting p53.

[0212] 206. The composition of aspect 205, wherein said induction of RNA interference targeting p53 is induced through administration of short interfering RNA.

[0213] 207. The composition of aspect 205, wherein said induction of RNA interference targeting p53 is induced through administration of short hairpin RNA.

[0214] 208. The composition of aspect 201, wherein said suppression of p53 expression is achieved through administration of antisense oligonucleotides targeting p53.

[0215] 209. The composition of aspect 208, wherein said antisense oligonucleotides induce cleavage of nucleic acids through activation of RNAse H.

[0216] 210. The composition of aspect 201, wherein said suppression of p53 expression is achieved through administration of ribozymes targeting p53.

[0217] 211. The composition of aspect 201, wherein said suppression of p53 expression is achieved through gene editing.

[0218] 212. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a liquid media containing ascorbic acid.

[0219] 213. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a liquid media containing transferrin.

[0220] 214. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a liquid media containing sodium bicarbonate.

[0221] 215. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a liquid media containing insulin.

[0222] 216. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a liquid media containing sodium selenite.

[0223] 217. The composition of aspect 200, wherein said reprogramming is induced by culture of said cells in a hypoxic environment.

[0224] 218. The composition of aspect 207, wherein said hypoxic environment comprises of oxygen levels low enough to induce activation of hypoxia inducible factor (HIF)-l.

[0225] 219. The composition of aspect 217, wherein said hypoxic environment is culture of said cells in an environment less than 21% oxygen.

[0226] 220. The composition of aspect 217, wherein said hypoxic environment is culture of said cells in an environment less than 15% oxygen.

[0227] 221. The composition of aspect 217, wherein said hypoxic environment is culture of said cells in an environment less than 10% oxygen.

[0228] 222. The composition of aspect 217, wherein said hypoxic environment is culture of said cells in an environment containing approximately 5% oxygen.

[0229] 223. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing a MAP Kinase inhibitor.

[0230] 224. The composition of aspect 200, wherein said MAP kinase inhibitor isPD0325901 /

[0231] 225. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing SB431542.

[0232] 226. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing CHIR99021.

[0233] 227. The composition of aspect 141, wherein said reprogramming is induced by culture of cells in a liquid media containing Y-27632.

[0234] 228. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing Y- thiazovivin.

[0235] 229. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing FGF-1.

[0236] 230. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing FGF-2.

[0237] 231. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing FGF-5.

[0238] 232. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing sodium borate.

[0239] 233. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing erythropoietin (EPO).

[0240] 234. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-3.

[0241] 235. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-6.

[0242] 236. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-8.

[0243] 237. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-10.

[0244] 238. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-18.

[0245] 239. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-20.

[0246] 240. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing interleukin-25.

[0247] 241. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing insulin-like growth factor-1 (IGF-1).

[0248] 242. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing dexamethasone.

[0249] 243. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing holo-transferrin.

[0250] 244. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing amino acids selected from a group comprising of Glycine, L-histidine, L-isoleucine, L-methionine, L-phenylalanine, L-proline, L-hydroxyproline, L-serine, L-threonine, L-tryptophan, and L-tyrosine, L-valine.

[0251] 245. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing vitamins and / or antioxidants selected from a group comprising of thiamine, reduced glutathione, ascorbic acid and 2-PO.sub.4.

[0252] 246. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing trace elements selected from a group comprising of: Ag.sup.+, Al.sup.3+, Ba.sup.2+, Cd.sup.2+, Co.sup.2+, Cr.sup.3+, Ge.sup.4+, Se.sup.4+, Br.sup.-, I. sup.-, F.sup.-, Mn.sup.2+, Si.sup.4+, V.sup.5+, MO.sup.6+, Ni.sup.2+, Rb.sup.+, Sn.sup.2+, and Zr.sup.4+.

[0253] 247. The composition of aspect 200, wherein said reprogramming is induced by culture of cells in a liquid media containing cytoplasm of an undifferentiated cell.

[0254] 248. The composition of aspect 247, wherein said cell being reprogrammed has its membrane temporarily permeabilized.

[0255] 249. The composition of 248, wherein said temporary permeabilization allows for entry of cytoplasm of undifferentiated cell into cytoplasm of said cell to be reprogrammed.

[0256] 250. The composition of aspect 247, wherein said undifferentiated cell is syngeneic with the cell whose reprogramming is desired.

[0257] 251. The composition of aspect 247, wherein said undifferentiated cell is allogeneic with the cell whose reprogramming is desired.

[0258] 252. The composition of aspect 247, wherein said undifferentiated cell is xenogeneic with the cell whose reprogramming is desired.

[0259] 253. The composition of aspect 248, wherein said permeabilization is mediated by electroporation.

[0260] 254. The composition of aspect 248, wherein said permeabilization is mediated by Streptolysin O treatment.

[0261] 255. The composition of aspect 248, wherein said permeabilization is mediated by transient treatment with complement membrane attack complex.

[0262] 256. The composition of aspect 248, wherein said permeabilization is mediated by transient treatment with perforin.

[0263] 257. The composition of aspect 248, wherein said permeabilization is mediated by transient treatment with granzyme.

[0264] 258. The composition of aspect 247, wherein said undifferentiated cell is an oocyte.

[0265] 259. The composition of aspect 258, wherein said oocyte is programmed to be at G0 / G1 of cell cycle.

[0266] 260. The composition of aspect 259, wherein said programming to be at G0 / G1 of cell cycle is accomplished by exposure to mitomycin C.

[0267] 261. The composition of aspect 259, wherein said programming to be at G0 / G1 of cell cycle is accomplished by exposure to serum starvation.

[0268] 262. The composition of aspect 247, wherein said undifferentiated cell is an inducible pluripotent stem cell.

[0269] 263. The composition of aspect 247, wherein said undifferentiated cell is a parthenogenic derived stem cell.

[0270] 264. The composition of aspect 247, wherein said undifferentiated cell is an embryonic stem cell.

[0271] 265. The composition of aspect 247, wherein said undifferentiated cell is a somatic cell nuclear transfer derived stem cell.

[0272] 267. The composition of aspect 247, wherein said undifferentiated cell is a cytoplasmically reprogrammed stem cell.

[0273] 268. The composition of aspect 247, wherein said undifferentiated cell is a cell obtained by fusion of an adult cell with a pluripotent stem cell.

[0274] 269. The composition of aspect 268, wherein said fusion is accomplished by the use of polyethylene glycol.

[0275] 270. The composition of aspect 268, wherein said fusion is accomplished by the use of electrically mediated fusion.

[0276] 271. A method for generating T regulatory cells comprising the steps of: a) obtaining a population of naive T cells; b) contacting said naive T cells with thecomposition of aspect 200 in a manner capable of eliciting immune modulation; and c) providing conditions so as to enable differentiation of naive T cells into T regulatory cells.

[0277] Til. The method of aspect 271, wherein said naive T cells are CD4 T cells.

[0278] 273. The method of aspect 271, wherein said naive T cells are CD8 T cells.

[0279] 274. The method of aspect 271, wherein said naive T cells are CD45RO T cells.

[0280] 275. The method of aspect 271, wherein said naive T cells are antigenically naive.

[0281] 276. The method of aspect 271, wherein said naive T cells express IL-2 receptor alpha chain.

[0282] TH. The method of aspect 271, wherein said cells of composition of aspect 1 are capable of immune modulation have been cultured in interferon gamma.

[0283] 278. The method of aspect 271, wherein said wherein said cells of composition of aspect 1 are capable of immune modulation have been cultured in interferon gamma

[0284] 279. The method of aspect 271, wherein said wherein said cells of composition of aspect 1 are capable of immune modulation have been cultured under hypoxic conditions.

[0285] 280. The method of aspect 271, wherein said wherein said cells of composition of aspect 1 are capable of immune modulation have been transfected with cytoplasm from immature dendritic cells.

[0286] 281. The method of aspect 280, wherein said immature dendritic cells lack substantial expression of CD40.

[0287] 282. The method of aspect 280, wherein said immature dendritic cells lack substantial expression of CD80.

[0288] 283. The method of aspect 280, wherein said immature dendritic cells lack substantial expression of CD86.

[0289] 284. The method of aspect 280, wherein said immature dendritic cells lack substantial expression of HLA II.

[0290] 285. The method of aspect 280, wherein said immature dendritic cells possessPD-1L.

[0291] 286. The method of aspect 280, wherein said immature dendritic cells possessILT-3.

[0292] 287. The method of aspect 280, wherein said immature dendritic cells secrete IL-10.

[0293] 288. The method of aspect 280, wherein said immature dendritic cells are derived from a cell line.

[0294] 289. The method of aspect 280, wherein said immature dendritic cells are derived from primary donors.

[0295] 290. The method of aspect 271, wherein said wherein said cells of composition of aspect 1 are capable of immune modulation have been cultured in platelet rich plasma.

[0296] 291. The method of aspect 271, wherein said wherein said cells of composition of aspect 1 are capable of immune modulation have been genetically modified to express an immune suppressive protein.

[0297] 292. The method of aspect 291, wherein said immune suppressive protein is IL-10.

[0298] 293. The method of aspect 291, wherein said immune suppressive protein isTGF-beta.

[0299] 294. The method of aspect 291, wherein said immune suppressive protein is IL-32.

[0300] 295. The method of aspect 291, wherein said immune suppressive protein is IL-35.

[0301] 296. The method of aspect 291, wherein said immune suppressive protein is IL-12p40 homodimers.

[0302] 297. The method of aspect 291, wherein said immune suppressive protein isHLA-G.

[0303] 298. The method of aspect 291, wherein said immune suppressive protein is ILT-3.

[0304] 299. The method of aspect 291, wherein said immune suppressive protein is indolamide 2,3 deoxygenase.

[0305] 300. The method of aspect 291, wherein said immune suppressive protein is indolamide 2,3 deoxygenase.

[0306] 301. A method of treating an inflammatory condition comprising the steps of: a) obtaining an pluripotent stem cell derived mesenchymal stem cell population; b) culturing said pluripotent stem cell derived mesenchymal stem cell population in conditions to allow for augmentation of an immune modulating effect; and c) administering said cell population into a patient in need of treatment.

[0307] 302. The method of aspect 301, wherein said inflammatory condition is an autoimmune condition.

[0308] 303. The method of aspect 302, wherein said autoimmune condition is a state in which immune cells of the patient recognize and attack tissue of said patient.

[0309] 304. The method of aspect 303, wherein said immune cells are T cells.

[0310] 305. The method of aspect 303, wherein said immune cells are B cells.

[0311] 306. The method of aspect 303, wherein said immune cells are NK cells.

[0312] 307. The method of aspect 301, wherein said inflammatory condition is characterized by increased production of inflammatory cytokines as compared to an age matched patient not suffering from said inflammatory condition.

[0313] 308. The method of aspect 307, wherein said inflammatory cytokine is TNF- alpha.

[0314] 309. The method of aspect 307, wherein said inflammatory cytokine is IL-1.

[0315] 310. The method of aspect 307, wherein said inflammatory cytokine is IL-6.

[0316] 311. The method of aspect 307, wherein said inflammatory cytokine is IL-11.

[0317] 312. The method of aspect 307, wherein said inflammatory cytokine is IL-12.

[0318] 313. The method of aspect 307, wherein said inflammatory cytokine is IL-17.

[0319] 314. The method of aspect 307, wherein said inflammatory cytokine is IL-18.

[0320] 315. The method of aspect 307, wherein said inflammatory cytokine is IL-21.

[0321] 316. The method of aspect 307, wherein said inflammatory cytokine is IL-33.

[0322] 317. The method of aspect 301, wherein said inflammatory condition is characterized by increased activation of the complement system as compared to an age matched patient not suffering from said inflammatory condition.

[0323] 318. The method of aspect 307, wherein said pluripotent stem cell populations are cultured in interferon gamma at a concentration and duration sufficient to induce anti-inflammatory properties of said pluripotent stem cell.

[0324] 319. The method of aspect 318, wherein said anti-inflammatory properties are selected from a group consisting of: a) suppression of ongoing mixed lymphocyte reaction; b) suppression of inflammatory cytokine production; and c) stimulation of T regulatory cells.

[0325] 320. The method of aspect 318, wherein said pluripotent stem cell populations are cultured in the presence of interferon gamma at a concentration of 1-100 lU / ml.

[0326] 321. The method of aspect 320, wherein said interferon gamma is used to treat pluripotent stem cells at a concentration of 10-75 lU / ml.

[0327] 322. The method of aspect 321, wherein said interferon gamma is used to treat pluripotent stem cells at a concentration of 25-50 lU / ml.

[0328] 323. The method of aspect 318, wherein said interferon gamma is used to treat pluripotent stem cells for a period of time ranging from 1 hour to 14 days.

[0329] 324. The method of aspect 318, wherein said interferon gamma is used to treat pluripotent stem cells for a period of time ranging from 1 to 7 days.

[0330] 325. The method of aspect 318, wherein said interferon gamma is used to treat pluripotent stem cells for a period of time ranging from 1 to 7 days.

[0331] 326. The method of aspect 318, wherein said biological response modifier is platelet rich plasma.

[0332] 327. The method of aspect 301, wherein platelet rich plasma used to treat pluripotent stem cell at a concentration of 1-50% volume by volume in tissue culture media in which said pluripotent stem cell are cultured.

[0333] 328. The method of aspect 327, wherein said platelet rich plasma used to treat pluripotent stem cells at a concentration of 5-20% volume by volume in tissue culture media in which said pluripotent cells are cultured.

[0334] 329. The method of aspect 327, wherein said platelet rich plasma used to treat pluripotent stem cells at a concentration of 5-10% volume by volume in tissue culture media in which said pluripotent cells are cultured.

[0335] 330. The method of aspect 327, wherein said platelet rich plasma is used to treat umbilical cord stem cells for a period of time ranging from 1 hour to 14 days.

[0336] 331. The method of aspect 327, wherein said platelet rich plasma is used to treat umbilical cord stem cells for a period of time ranging from 1 to 7 days.

[0337] 332. The method of aspect 327, wherein said platelet rich plasma is used to treat umbilical cord stem cells for a period of time ranging from 1 to 7 days.

[0338] 333. The method of aspect 301, wherein said umbilical cord stem cells are cultured in a media selected from a group comprising of: a) Roswell Park Memorial Institute (RPMI-1640); b) Dublecco's Modified Essential Media (DMEM), c) Eagle's Modified Essential Media (EMEM), d) Optimem, and e) Iscove's Media.

[0339] 334. The method of aspect 301, wherein said umbilical cord stem cells are selected for expression of CD73.

[0340] 335. The method of aspect 334, wherein selection for CD73 is performed during isolation of umbilical cord mononuclear cells from said umbilical cord tissue.

[0341] 336. The method of aspect 335, wherein said isolation of said umbilical cord mononuclear cells is performed from said umbilical cord tissue by use of enzymatic digestion.

[0342] 337. The method of aspect 335, wherein said isolation of said umbilical cord mononuclear cells is performed from said umbilical cord tissue by use of mechanical dissociation.

[0343] 338. The method of aspect 335, wherein said isolation of said umbilical cord mononuclear cells is performed from said umbilical cord tissue by use of mechanical dissociation and enzymatic digestion.

[0344] 339. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of interleukin-7 receptor.

[0345] 340. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of interleukin-3 receptor.

[0346] 341. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of Receptor for Advanced Glycation End Products (RAGE).

[0347] 342. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TNF-alpha receptor p55.

[0348] 343. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TNF-alpha receptor p75.

[0349] 344. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of stem cell factor receptor.

[0350] 345. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of GM-CSF receptor alpha.

[0351] 346. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of VPR-Binding Protein.

[0352] 347. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of transferrin receptor.

[0353] 348. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of Tmc5 protein.

[0354] 349. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-9.

[0355] 350. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of zinc transporter 9.

[0356] 351. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of seminal vesicle antigen-like 3.

[0357] 352. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of sarcoma antigen NY-SAR-41.

[0358] 353. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of cell surface vimentin.

[0359] 354. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of fibrosin-1.

[0360] 355. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of IL-1 receptor.

[0361] 356. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of IL-3 receptor.

[0362] 357. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of IL-6 receptor.

[0363] 358. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of HGF receptor.

[0364] 359. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of thrombopoietin receptor.

[0365] 360. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of prolactin receptor.

[0366] 361. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of IGF-1 receptor.

[0367] 362. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of PDGF-BB receptor.

[0368] 363. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of angiopoietin receptor.

[0369] 364. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of VEGF receptor.

[0370] 365. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-2.

[0371] 366. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-3.

[0372] 367. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-4.

[0373] 368. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-5.

[0374] 369. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-7.

[0375] 370. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of TLR-8.

[0376] 371. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of oxysterol-binding protein 1.

[0377] 372. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of nesprin-2.

[0378] 373. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of myomesin-3.

[0379] 374. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of mucin-2.

[0380] 375. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of FRASl-related extracellular matrix protein 3.

[0381] 376. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of C-C chemokine receptor type 10.

[0382] 377. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of CXCR4.

[0383] 378. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of CCR5

[0384] 379. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of cartilage intermediate layer protein 2.

[0385] 380. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of H LA-DR.

[0386] 381. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of oxytocin receptor.

[0387] 382. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of CD77.

[0388] 383. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of CD56.

[0389] 384. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of poliovirus receptor.

[0390] 385. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of plexin A2.

[0391] 386. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of plexin A4.

[0392] 387. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of HLA-G.

[0393] 388. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of membrane bound TGF-beta.

[0394] 389. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of membrane bound TNF-alpha.

[0395] 390. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of plasticity-related protein 2.

[0396] 391. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of BDNF receptor.

[0397] 392. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of occludin.

[0398] 393. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of neuronal pentraxin receptor.

[0399] 394. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of neuropilin-1.

[0400] 395. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of netrin 2-like.

[0401] 396. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of mucolipin 1.

[0402] 397. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of MEGF10 protein.

[0403] 398. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of mannose binding lectin (A).

[0404] 399. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of Leukemia Inhibitory Factor Receptor.

[0405] 400. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of lipocalin 3.

[0406] 401. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of lipocalin 12.

[0407] 402. The method of aspect 335, wherein said CD73 expressing cells are further selected for expression of lipocalin 13.

[0408] 403. A method of treating cancer comprising: a) selecting an umbilical cord derived mononuclear cell population, ideally of mesenchymal phenotype; b) increasing ability said cells to home to tumor microenvironment; c) increasing ability of cells to inhibit cancer; and d) administering said cells to a patient in need of treatment.

[0409] 404. The method of aspect 403, wherein said increased ability of said cell to home to tumors comprises incubation of said cell in a hypoxic environment from 0.1% oxygen to 10% oxygen for a period of 30 minutes to 3 days.

[0410] 405. The method of aspect 404, wherein said cell is cultured at 3% oxygen for24 hours.

[0411] 406. The method of aspect 403, wherein said increased ability of said cell to inhibit cancer is accomplished by infection of said cell with an oncolytic virus.

[0412] 407. The method of aspect 406, wherein said oncolytic virus is selected from a group comprising of: a) vaccinia virus; b) reovirus; c) Newcastle Disease Virus; d) herpes virus; e) parvovirus; f) measles virus; g) vesicular stomatitis virus (VSV); h) adenovirus; i) poliovirus; j) a poxvirus; k) coxsackie virus (CXV); and I) Seneca Valley virus (SVV).

[0413] 408. The cell of aspect 407, wherein the vaccinia virus is selected from among aLister strain, Western Reserve (WR) strain, Copenhagen (Cop) strain, Bern strain, Paris strain, Tashkent strain, Tian Tan strain, Wyeth strain (DRYVAX), IHD-J strain, IHD-W strain, Brighton strain, Ankara strain, CVA382 strain, Dairen I strain, LC16m8 strain, LC16M0 strain, modified vaccinia Ankara (MVA) strain, ACAM strain, WR 65-16 strain, Connaught strain, New York City Board of Health (NYCBH) strain, EM-63 strain, NYVAC strain, Lister strain LIVP, JX-594 strain, GL-ONC1 strain, a vvDD TK mutant strain with deletions in VGF and TK, ACAM2000, and ACAM1000

[0414] 409. The method of aspect 403, wherein said increased ability of cells to inhibit cancer is endowed by transfection of said cells with a cancer-inhibitory gene.

[0415] 410. The method of aspect 409, wherein said cancer inhibitory gene is under control of an inducible promoter.

[0416] 411. The method of aspect 410, wherein said inducible promoter is a rheoswitch.

[0417] 412. The method of aspect 409, wherein said cancer-inhibitory gene is TNF- alpha.

[0418] 413. The method of aspect 409, wherein said cancer-inhibitory gene is TRAIL.

[0419] 414. The method of aspect 409, wherein said cancer-inhibitory gene is a suicide gene.

[0420] 415. The method of aspect 409, wherein said cancer-inhibitory gene is thymidylate synthesase.

[0421] 416. The method of aspect 403, wherein said increased ability of cells to inhibit cancer is endowed by transfection of said cells with an immune stimulatory gene.

[0422] 417. The method of aspect 416, wherein said immune stimulatory gene is IL-2.

[0423] 418. The method of aspect 416, wherein said immune stimulatory gene is IL-15.

[0424] 419. The method of aspect 416, wherein said immune stimulatory gene is IL-7.

[0425] 420. The method of aspect 416, wherein said immune stimulatory gene is IL-12.

[0426] 421. The method of aspect 416, wherein said immune stimulatory gene is IL-15.

[0427] 422. The method of aspect 416, wherein said immune stimulatory gene is IL-22.

[0428] 423. The method of aspect 416, wherein said immune stimulatory gene is IL-18.

[0429] 424. The method of aspect 416, wherein said immune stimulatory gene is IL-27.

[0430] 425. The method of aspect 416, wherein said immune stimulatory gene is a bispecific antibody.

[0431] 426. The method of aspect 406, wherein said infection with an oncolytic virus is performed in an umbilical cord derived mesenchymal stem cell that is treated with hypoxia in a manner sufficient to induce translocation of HIF-1 alpha.

[0432] 427. The method of aspect 426, wherein said cell is engineered to express an oncogene.

[0433] 428. The method of aspect 427, wherein said oncogene is selected from a group comprising of: ABCB1, ABCG2, ABI1, ABL1, ABL2, ACKR3, ACSL3, ACSL6, ACVR1B, ACVR2A, AFF1, AFF3, AFF4, AKAP9, AKT1, AKT2, AKT3, ALDH1A1, ALDH2, ALK, AMER1, ANGPT1, ANGPT2, ANKRD23, APC, AR, ARAF, AREG, ARFRP1, ARHGAP26, ARHGEF12, ARID1A, ARID1B, ARID2, ARNT, ASPSCR1, ASXL1, ATF1, ATIC, ATM, ATP1A1, ATP2B3, ATR, ATRX, AURKA, AURKB, AXIN1, AXL, BAP1, BARD1, BBC3, BCL10, BCL11A, BCL11B, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL3, BCL6, BCL7A, BCL9, BCOR, BCORL1, BCR, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD3, BRD4, BRINP3, BRIP1, BTG1, BTG2, BTK, BUB1B, Cllorf30, C15orf65, C2orf44, CA6, CACNA1D, CALR, CAMTAI, CANT1, CARD11, CARS, CASC5, CASP8, CBFA2T3, CBFB, CBL, CBLB, CBLC, CCDC6, CCNB1IP1, CCND1, CCND2, CCND3, CCNE1, CD19, CD22, CD274, CD38, CD4, CD70, CD74, CD79A, CD79B, CD83, CDC73, CDH1, CDH11, CDK12, CDK4, CDK6, CDK7, CDK8, CDK9, CDKN1A, CDKN1B, CDKN2A, CDKN2B, CDKN2C, CDX2, CEBPA, CHCHD7, CHD2, CHD4, CHEK1, CHEK2, CHIC2, 1CHN1, CHORDCI, CIC, CIITA, CLP1, CLTC, CLTCL1, CNBP, CNOT3, CNTRL, COL1A1, COPB1, COX6C, CRBN, CREB1, CREB3L1, CREB3L2, CREBBP, CRKL, CRLF2, CRTC1, CRTC3, CSF1R, CSF3R, CTCF, CTLA4, CTNNA1, CTNNB1, CUL3, CXCR4, CYLD, CYP17A1, CYP2D6, DAXX, DDB2, DDIT3, DDR1, DDR2, DDX10, DDX3X, DDX5, DDX6, DEK, DICER1, DIS3, DLL4, DNM2, DNMT1, DNMT3A, DOTH, DPYD, DUSP4, DUSP6, EBF1, ECT2L, EDNRB, EED, EGFR, EIF4A2, ELF4, ELK4, ELL, ELN, EML4, EP300, EPHA3, EPHA5, EPHA7, EPHA8, EPHB1, EPHB2, EPHB4, EPS15, ERBB2, ERBB3, ERBB4, ERC1, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, EREG, ERG, ERN1, ERRFI1, ESRI, ETV1, ETV4, ETV5, ETV6, EWSR1, EXT1, EXT2, EZH2, EZR, FAF1, FAIM3, FAM46C, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCL, FAS, FAT1, FBXO11, FBXW7, FCRL4, FEV, FGF1O, FGF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF6, FGFR1, FGFR1OP, FGFR2, FGFR3, FGFR4, FH, FHIT, FIP1L1, FKBP1A, FLCN, FLU, FLT1, FLT3, FLT4, FNBP1, FOXA1, FOXL2, FOXO1, FOXO3, FOXO4, FOXP1, FRS2, FSTL3, FUBP1, FUS, GABRA6, GAS7, GATA1, GATA2, GATA3, GATA4, GATA6, GID4, GUI, GM PS, GNA11, GNA12, GNA13, GNAQ, GNAS, GNRH1, GOLGA5, GOPC, GPC3, GPHN, GPR124, GRIN2A, GRM3, GSK3B, GUCY2C, H3F3A, H3F3B, HCK, HDAC1, HERPUD1, HEY1, HGF, HIP1, HIST1H1E, HIST1H3B, HIST1H4I, HLF, HMGA1, HMGA2, HMGN2P46, HNF1A, HNMT, HNRNPA2B1, HNRNPK, HOOK3, HOXA11, HOXA13, HOXA9, HOXC11, HOXC13, HOXD11, HOXD13, HRAS, HSD3B1, HSP90AA1, HSP90AB1, IAPP, ID3, IDH1, IDH2, IGF1R, IGF2, IKBKE, IKZF1, IL2, IL21R, IL3RA, IL6, IL6ST, IL7R, INHBA, INPP4B, IRF2, IRF4, IRS2, ITGAV, ITGB1, ITK, ITPKB, JAK1, JAK2, JAK3, JAZF1, JUN, KAT6A, KAT6B, KCNJ5, KDM1A, KDM5A, KDM5C, KDM6A, KDR, KDSR, KEAP1, KEL, KIAA1549, KIF5B, KIR3DL1, KIT, KLF4, KLHL6, KLK2, KMT2A, KMT2C, KMT2D, KRAS, KTN1, LASPI, LCK, LCP1, LGALS3, LGR5, LHFP, LIFR, LMO1, LMO2, LOXL2, LPP, LRIG3, LRP1B, LUC7L2, LYL1, LYN, LZTR1, MAF, MAFB, MAGED1, MAGI2, MALT1, MAM L2, MAP2K1, MAP2K2, MAP2K4, MAP3K1, MAPK1, MAPK11, MAX, MCL1, MDM2, MDM4, MDS2, MECOM, M ED12, M EF2B, MEN1, MET, MITF, M KI67, MKL1, M LF1, MLH1, MLLT1, MLLT10, MLLT11, MLLT3, MLLT4, MLLT6, MM P9, M N1, MNX1, MPL, MRE11A, MS4A1, MSH2, MSH6, MSI2, MSN, MST1R, MTCP1, MTF2, MTOR, MUC1, MUC16, M UTYH, MYB, MYC, MYCL, MYCN, MYD88, MYH11, MYH9, NACA, NAE1, NBN, NCKIPSD, NCOA1, NCOA2, NCOA4, NDRG1, NF1, NF2, NFE2L2, NFIB, NFKB2, NFKBIA, NIN, NKX2-1, NONO, NOTCH1, NOTCH2, NOTCH3, NPM1, NR4A3, NRAS, NSD1, NT5C2, NTRK1, NTRK2, NTRK3, NUMA1, NUP214, NUP93, NUP98, NUTM1, NUTM2B, OLIG2, OMD, P2RY8, PAFAH1B2, PAK3, PALB2, PARK2, PARP1, PATZ1, PAX3, PAX5, PAX7, PAX8, PBRM 1, PBX1, PCM 1, PCSK7, PDCD1, PDCD1LG2, PDE4DIP, PDGFB, PDGFRA, PDGFRB, PDK1, PECAM1, PERI, PHF6, PHOX2B, PICALM, PIK3C2B, PIK3CA,PIK3CB, PIK3CD, PIK3CG, PIK3R1, PIK3R2, PIM1, PLAG1, PLCG2, PML, PMS1, PMS2, POLDI, POLE, POTI, POU2AF1, POU5F1, PPARG, PPP2R1A, PRCC, PRDM1, PRDM16, PREX2, PRF1, PRKAR1A, PRKCI, PRKDC, PRLR, PRPF40B, PRRT2, PRRX1, PRSS8, PSIP1, PSMD4, PTBP1, PTCHI, PTEN, PTK2, PTPN11, PTPRC, PTPRD, QKI, RABEP1, RAC1, RAD21, RAD50, RAD51, RAD51B, RAD51C, RAD51D, RAFI, RALGDS, RANBP17, RANBP2, RAP1GDS1, RARA, R131, RBM 10, RBM15, RCOR1, RECQL4, REL, RELN, RET, RHOA, RHOH, RICTOR, RIPK1, RMI2, RNF213, RNF43, ROS1, RPL1O, RPL22, RPL5, RPN1, RPS6KB1, RPTOR, RUNX1, RUNX1T1, S1PR2, SAMHD1, SBDS, SDC4, SDHA, SDHAF2, SDHB, SDHC, SDHD, SEPT5, SEPT6, SEPT9, SET, SETBP1, SETD2, SF1, SF3A1, SF3B1, SF3B2, SFPQ, SGK1, SH2B3, SH3GL1, SLAM F7, SLC34A2, SLC45A3, SLIT2, SMAD2, SMAD3, SMAD4, SMARCA4, SMARCB1, SMARCE1, SMC1A, SMC3, SMO, SNCAIP, SNX29, SOCS1, SOXIO, SOX11, SOX2, SOX9, SPECC1, SPEN, SPOP, SPTA1, SRC, SRGAP3, SRSF2, SRSF3, SS18, SS18L1, SSX1, STAG2, STAT3, STAT4, STAT5B, STEAP1, STIL, STK11, SUFU, SUZ12, SYK, TAF1, TAF15, TALI, TAL2, TBL1XR1, TBX3, TCEA1, TCF12, TCF3, TCF7L2, TCL1A, TEK, TERC, TERT, TET1, TET2, TFE3, TFEB, TFG, TFPT, TFRC, TGFB1, TGFBR2, THRAP3, TIMP1, TJP1, TLX1, TLX3, TM7SF2, TMPRSS2, TNFAIP3, TNFRSF14, TNFRSF17, TNFRSF18, TNFRSF9, TNFSF11, TOPI, TOP2A, TP53, TP63, TPBG, TPM3, TPM4, TPR, TRAF2, TRAF3, TRAF3IP3, TRAF7, TRIM26, TRIM27, TRIM33, TRIP11, TRRAP, TSC1, TSC2, TSHR, TTK, TTL, TYMS, U2AF1, U2AF2, UBA1, UBR5, USP6, VEGFA, VEGFB, VHL, VPS51, VTI1A, WAS, WEE1, WHSCI, WHSC1L1, WIFI, WISP3, WNT11, WNT2B, WNT3, WNT3A, WNT4, WNT5A, WNT6, WNT7B, WRN, WT1, WWTR1, XBP1, XPA, XPC, XPO1, YWHAE, YWHAZ, ZAK, ZBTB16, ZBTB2, ZMYM2, ZMYM3, ZNF217, ZNF331, ZNF384, ZNF521, ZNF703 and ZRSR2

[0434] 429. The method of aspect 426, wherein said cell is engineered to express a tumor suppressor gene.

[0435] 430. The method of aspect 429, wherein said tumor suppressor gene is selected from a group comprising of: P53. RBI, WT1, NF1, NF2, APC, TSC1, TSC2, DPC4, DCC, BRCA1, BRCA2, PTEN, STK11, MSH2, MLH1, CDH1, VHL, CDKN2A, PTCH, MEN1.

[0436] 432. The method of aspect 426, wherein said cell is gene edited, and / or treated with an agent or plurality of agents which induce RNA interference, and / or treated with an agent or plurality of agents which induce RNAse U in order to reduce or remove expression of a checkpoint molecule.

[0437] 433. The method of aspect 432, wherein said checkpoint molecules are selected from a group comprising of: PD1 (also called PDCD1 or CD279); PD-L1 (also called B7-H1 or CD274); PD-L2 (also called B7-DC or CD273); CTLA-4 (also called CD152); B7-H3 (alsocalled CD276); B7-H4 (also called B7S1 or B7x); CD66a (CEACAM1); VISTA (also called B7- H5 or GI24); BTLA; CD160; LAGS (also called CD223 or Lymphocyte activation gene 3); Indoleamine 2,3-dioxygenase (also called IDO); Galectin-9 (also called LGALS9); TIM-3 (also called HAVCR2); 2B4 (also called CD244); SIRP alpha (also called CD172a); CD39; CD47; CD48 (also called SLAMF2); A2AR; KIRs; and TIGIT (also called VSTM3).

[0438] 434. The method of aspect 406, wherein said cell is incubated with the virus for at least 16 hours or at least 20 hours or at least 24 hours.

[0439] 435. The method of aspect 406, wherein said cell is incubated with the virus for up to 48 hours.

[0440] 436. The method of aspect 406, wherein said cell is infected with virus at a MOI less than or equal to 0.8.

[0441] 437. The method of aspect 406, wherein said cell is infected with virus at a MOI less than or equal to 0.5.

[0442] 438. The method of aspect 403 wherein said cell isolated from said Wharton'sJelly express a marker selected from a group of markers comprising: CD56, CD57, CD144, CD105, and CD31.

[0443] 439. The composition of aspect 438, wherein said cells are obtained from placenta perivascular tissue as a substitute for Wharton's Jelly.

[0444] 440. The composition of aspect 439, wherein said cells are s isolated from fetal vascular lobules of a hemochorial placenta.

[0445] 441. The composition of aspect 439, wherein said cells are isolated by: dissociating fetal vascular lobules from a hemochorial placenta; digesting the dissociated fetal vascular lobes with an enzymatic mixture or by mechanical means; applying a filtration means to said dissociated lobes in order to remove particulates; obtaining mononuclear cells; plating said mononuclear cells in a substrate allowing for growth of said mononuclear cells to confluency; detaching the confluent cells from the plate; and isolating for expression of CD144 and substantially lack of expression of CD45, optionally one or more steps are performed in the presence of hypoxia, wherein hypoxia is sufficient to induce translocation of HIF-1 alpha.

[0446] 442. The method of aspect 441, wherein dissociation of fetal vascular lobes is accomplishing by incubation with a mixture of about 2% collagenase, about 0.25% trypsin and about 0.1% DNAse in tissue culture medium.

[0447] 443. The method of aspect 439, wherein dissociation of fetal vascular lobes is accomplishing by incubation with a mixture of about 2% collagenase, about 0.25% trypsin and about 0.1% DNAse in tissue culture medium.

[0448] 444. The composition of aspect 438, wherein cells isolated are comprised of adherent cells expressing the marker CD73 but substantially lacking CD105.

[0449] 445. The composition of aspect 438, wherein cells isolated are comprised of adherent cells expressing the marker CD73 and CD105 but lacking in CD90.

[0450] 446. The composition of aspect 438, wherein said regenerative adjuvant is an anti-inflammatory cytokine.

[0451] 447. The composition of aspect 446, wherein said anti-inflammatory cytokine is selected from a group comprising of IL-4, IL-10, IL-13, IL-20, IL-22 and IL-35.

[0452] 448. The composition of aspect 446, wherein said anti-inflammatory cytokine isTGF-beta.

[0453] 449. The composition of aspect 446, wherein said anti-inflammatory cytokine isPGE-2.

[0454] 450. The composition of aspect 446, wherein said anti-inflammatory cytokine isVEGF.

[0455] 451. The composition of aspect 438, wherein said composition is composed of mesenchymal stem cells, and wherein said first regenerative adjuvant is hypoxia.

[0456] 452. The composition of aspect 438, wherein said mesenchymal stem cells possess one or more markers selected from a group comprising of: a) CDllb; b) CDllc; c) CD20; d) CD56; e) CD57 f) CD73; g) CD90; h) CD105; i) membrane bound TGF-beta; and j) neuropilin.

[0457] 453. The composition of aspect 438, wherein said composition is capable of inhibiting T cell mediated immune responses.

[0458] 454. The method of aspect 453, wherein said T cell mediated immune responses comprise of Thl cell production of cytokines.

[0459] 455. The method of aspect 403, wherein said cancers are selected from a group of malignancies comprising of: acute lymphoblastic leukemia; acute myeloid leukemia; adrenocortical carcinoma; AIDS-related cancer; AIDS-related lymphoma; anal cancer; appendix cancer; astrocytomas; atypical teratoid / rhabdoid tumor; basal cell carcinoma; bladder cancer; brain stem glioma; brain tumor, brain stem glioma, central nervous system atypical teratoid / rhabdoid tumor, central nervous system embryonal tumors, astrocytomas, craniopharyngioma, ependymoblastoma, ependymoma,medulloblastoma, medulloepithelioma, pineal parenchymal tumors of intermediate differentiation, supratentorial primitive neuroectodermal tumors and pineoblastoma; breast cancer; bronchial tumors; Burkitt lymphoma; cancer of unknown primary site (CUP); carcinoid tumor; carcinoma of unknown primary site; central nervous system atypical teratoid / rhabdoid tumor; central nervous system embryonal tumors; cervical cancer; childhood cancers; chordoma; chronic lymphocytic leukemia; chronic myelogenous leukemia; chronic myeloproliferative disorders; colon cancer; colorectal cancer; craniopharyngioma; cutaneous T-cell lymphoma; endocrine pancreas islet cell tumors; endometrial cancer; ependymoblastoma; ependymoma; esophageal cancer; esthesioneuroblastoma; Ewing sarcoma; extracranial germ cell tumor; extragonadal germ cell tumor; extrahepatic bile duct cancer; gallbladder cancer; gastric (stomach) cancer; gastrointestinal carcinoid tumor; gastrointestinal stromal cell tumor; gastrointestinal stromal tumor (GIST); gestational trophoblastic tumor; glioma; hairy cell leukemia; head and neck cancer; heart cancer; Hodgkin lymphoma; hypopharyngeal cancer; intraocular melanoma; islet cell tumors; Kaposi sarcoma; kidney cancer;Langerhans cell histiocytosis; laryngeal cancer; lip cancer; liver cancer; malignant fibrous histiocytoma bone cancer; medulloblastoma; medulloepithelioma; melanoma; Merkel cell carcinoma; Merkel cell skin carcinoma; mesothelioma; metastatic squamous neck cancer with occult primary; mouth cancer; multiple endocrine neoplasia syndromes; multiple myeloma; multiple myeloma / plasma cell neoplasm; mycosis fungoides; myelodysplastic syndromes; myeloproliferative neoplasms; nasal cavity cancer; nasopharyngeal cancer; neuroblastoma; Non-Hodgkin lymphoma; nonmelanoma skin cancer; non-small cell lung cancer; oral cancer; oral cavity cancer; oropharyngeal cancer; osteosarcoma; other brain and spinal cord tumors; ovarian cancer; ovarian epithelial cancer; ovarian germ cell tumor; ovarian low malignant potential tumor; pancreatic cancer; papillomatosis; paranasal sinus cancer; parathyroid cancer; pelvic cancer; penile cancer; pharyngeal cancer; pineal parenchymal tumors of intermediate differentiation; pineoblastoma; pituitary tumor; plasma cell neoplasm / multiple myeloma; pleuropulmonary blastoma; primary central nervous system (CNS) lymphoma; primary hepatocellular liver cancer; prostate cancer; rectal cancer; renal cancer; renal cell (kidney) cancer; renal cell cancer; respiratory tract cancer; retinoblastoma; rhabdomyosarcoma; salivary gland cancer; Sezary syndrome; small cell lung cancer; small intestine cancer; soft tissue sarcoma; squamous cell carcinoma; squamous neck cancer; stomach (gastric) cancer; supratentorial primitive neuroectodermal tumors; T-cell lymphoma; testicular cancer; throat cancer; thymic carcinoma; thymoma; thyroid cancer; transitional cell cancer; transitional cell cancer of the renal pelvis and ureter; trophoblastic tumor; ureter cancer; urethral cancer; uterine cancer; uterine sarcoma; vaginal cancer; vulvar cancer; Waldenstrom macroglobulinemia; or Wilm's tumor.

[0460] 456. A method of inducing immunological tolerance, and / or predisposing to immunological tolerance, comprising the steps of: a) obtaining a population of umbilical cord mononuclear cells; b) exposing said cells to conditions capable of triggering apoptosis; c) extracting said apoptotic bodies; and d) administering said apoptotic bodies to a patient in need of treatment at a concentration and frequency sufficient to induce and / or predispose to immune tolerance.

[0461] 457. The method of aspect 456, wherein said tolerance is a state of nonresponsiveness of an immune cell to a condition or plurality of conditions normally results in responsiveness of said immune cell.

[0462] 458. The method of aspect 457, wherein said immune cell is an adaptive or innate immune cell.

[0463] 459. The method of aspect 458, wherein said adaptive immune cell is a T cell.

[0464] 460. The method of aspect 458, wherein said adaptive immune cell is a B cell.

[0465] 461. The method of aspect 458, wherein said innate immune cell is a monocyte.

[0466] 462. The method of aspect 458, wherein said innate immune cell is a natural killer cell.

[0467] 463. The method of aspect 458, wherein said innate immune cell is a gamma delta T cell.

[0468] 464. The method of aspect 458, wherein said innate immune cell is a natural killer T cell.

[0469] 465. The method of aspect 458, wherein said innate immune cell is a neutrophil.

[0470] 466. The method of aspect 458, wherein said innate immune cell is a dendritic cell.

[0471] 467. The method of aspect 456, wherein immunological tolerance is associated with remission in an autoimmune condition.

[0472] 468. The method of aspect 456, wherein said cells are adherent cells.

[0473] 469. The method of aspect 456, wherein said cells are obtained by positive selection of umbilical cord blood mononuclear cells for CD56.

[0474] 470. The method of aspect 456, wherein said cells express CD90.

[0475] 471. The method of aspect 456, wherein said cells express vimentin.

[0476] 472. The method of aspect 456, wherein said cells express CD57.

[0477] 473. The method of aspect 459, wherein said non-responsiveness of said T cell comprises reduction of interleukin-2 production in response to stimulation via T cell receptor.

[0478] 474. The method of aspect 459, wherein said non-responsiveness of said T cell comprises reduction of interferon gamma production in response to stimulation via T cell receptor.

[0479] 475. The method of aspect 459, wherein said non-responsiveness of said T cell comprises reduction of proliferation in response to stimulation via T cell receptor.

[0480] 476. The method of aspect 459, wherein said non-responsiveness of said T cell comprises reduction of interleukin-4 production in response to stimulation via T cell receptor.

[0481] 477. The method of aspect 459, wherein said non-responsiveness of said T cell comprises enhanced need for costimulation in order to activate said T cell subsequent to stimulation via T cell receptor.

[0482] 478. The method of aspect 459, wherein said non-responsiveness of said T cell comprises enhanced need for costimulation in order to activate said T cell subsequent to stimulation via T cell receptor.

[0483] 479. The method of aspect 459, wherein said non-responsiveness comprises reduction in cytotoxicity subsequent to stimulation via T cell receptor.

[0484] 480. The method of aspect 460, wherein said non-responsiveness comprises reduction in antibody production subsequent to stimulation via B cell receptor.

[0485] 481. The method of aspect 461, wherein said non-responsiveness comprises reduction in monocyte phagocytic activity.

[0486] 482. The method of aspect 461, wherein said non-responsiveness comprises reduction in monocyte cytokine production activity.

[0487] 483. The method of aspect 482, wherein said cytokines are selected from a group comprising of: a) TNF-alpha; b) IL-1 beta; c) IL-6; d) IL-8; e) IL-12; f) IL-18; g) IL-17; h) IL-21; i) IL-23; j) IL-27; k) IL-33; and I) RANTES.

[0488] 484. The method of aspect 482, wherein said non-responsiveness comprises reduction of Ml polarization.

[0489] 485. The method of aspect 482, wherein said non-responsiveness comprises enhancement M2 polarization.

[0490] 486. The method of aspect 482, wherein said non-responsiveness comprises inhibition of maturation to macrophages.

[0491] 487. The method of aspect 482, wherein said non-responsiveness comprises inhibition of maturation to dendritic cells.

[0492] 488. The method of aspect 482, wherein said non-responsiveness comprises inhibition of antigen presentation.

[0493] 489. The method of aspect 462, wherein said non-responsiveness comprises inhibition of NK cytotoxicity.

[0494] 490. The method of aspect 462, wherein said non-responsiveness comprises inhibition of NK ability to induce dendritic cell maturation.

[0495] 491. The method of aspect 462, wherein dendritic cell maturation comprises augmented expression of costimulatory molecules.

[0496] 492. The method of aspect 491, wherein said costimulatory molecule is CD40.

[0497] 493. The method of aspect 491, wherein said costimulatory molecule is CD80.

[0498] 494. The method of aspect 491, wherein said costimulatory molecule is CD86.

[0499] 495. The method of aspect 491, wherein said costimulatory molecule is IL-12.

[0500] 496. The method of aspect 463, wherein said non-responsiveness comprises inhibition of IL-12 secretion by gamma delta T cells.

[0501] 497. The method of aspect 463, wherein said non-responsiveness comprises inhibition of interferon gamma secretion by gamma delta T cells.

[0502] 498. The method of aspect 463, wherein said non-responsiveness comprises inhibition of cytotoxicity by gamma delta T cells.

[0503] 499. The method of aspect 463, wherein said non-responsiveness comprises inhibition of dendritic cell maturation by gamma delta T cells.

[0504] 500. The method of aspect 464, wherein said non-responsiveness comprises inhibition of natural killer T cell production of IL-18.

[0505] 501. The method of aspect 465, wherein said non-responsiveness comprises inhibition of neutrophil production of reactive oxidative intermediaries.

[0506] 502. The method of aspect 465, wherein said non-responsiveness comprises inhibition of neutrophil production of DNA extracellular traps.

[0507] 503. The method of aspect 465, wherein said non-responsiveness comprises induction of neutrophil apoptosis.

[0508] 504. The method of aspect 465, wherein said extraction of apoptotic bodies is performed by means of a density gradient.

[0509] 505. The method of aspect 465, wherein said extraction of apoptotic bodies is performed by an affinity means.

[0510] 506. The method of aspect 465, wherein said affinity means comprises binding to annexin V.

[0511] 507. The method of aspect 456, wherein apoptosis is induced by irradiation.

[0512] 508. The method of aspect 507, wherein said irradiation is gamma irradiation.

[0513] 509. The method of aspect 507, wherein said irradiation is X-irradiation.

[0514] 510. The method of aspect 507, wherein said irradiation is bombardment with helium nuclei.

[0515] 511. The method of aspect 507, wherein said irradiation is beta irradiation.

[0516] 512. The method of aspect 507, wherein said irradiation is muon irradiation.

[0517] 513. The method of aspect 507, wherein said irradiation is exposure to ultraviolet irradiation.

[0518] 514. The method of aspect 513, wherein a photosensitizer is added.

[0519] 515. The method of aspect 514, wherein said photosensitizer is psoralen.

[0520] 516. The method of aspect 456, wherein apoptosis is induced by exposure to an agent stimulating oxidative stress.

[0521] 517. The method of aspect 516 wherein said agent stimulating oxidative stress is ozone.

[0522] 518. The method of aspect 516, wherein said agent stimulating oxidative stress is hydrogen peroxide.

[0523] 519. The method of aspect 516, wherein said agent stimulating oxidative stress is hydroxyl radical.

[0524] 520. The method of aspect 516, wherein said agent stimulating oxidative stress is hyperthermia.

[0525] 521. The method of aspect 516, wherein said agent stimulating oxidative stress is non-isotonic conditions.

[0526] 522. The method of aspect 456, wherein said apoptosis is induced by exposure to TNF-alpha.

[0527] 523. The method of aspect 456, wherein said apoptosis is induced by exposure to TRAIL.

[0528] 524. The method of aspect 456, wherein said apoptosis is induced by exposure to Fas Ligand.

[0529] 525. The method of aspect 456, wherein immunological tolerance is used to promote pregnancy in a patient suffering from recurrent spontaneous abortions.

[0530] 526. The method of aspect 456, wherein said immunological tolerance is used to allow for decreased immune suppressants in a patient having received an organ, tissue, or cellular transplant.

[0531] 527. The method of aspect 456, wherein said immunological tolerance is used to allow for use of no immune suppressants in a patient having received an organ, tissue, or cellular transplant.

[0532] 528. A method of stimulating angiogenesis comprising the steps of: a) obtaining one or more umbilical cord adherent cell populations; b) culturing said umbilical cord cell populations in a manner to allow for production of exosomes into culture media in which said cells are cultured in; c) extracting exosomes from said culture media; and d) administering said extracted exosomes into a patient in need of treatment.

[0533] 529. The method of aspect 528, wherein said umbilical cord cells are derived from a term pregnancy, wherein said donor providing said cells is either the patient to be treated (autologous) or said donor is different from the patient to be treated (allogeneic).

[0534] 530. The method of aspect 528, wherein said cells are cultured in a media allowing for cell proliferation.

[0535] 531. The method of aspect 530, wherein said media allowing for cell proliferation contains one or more factors known to be mitogenic for cells.

[0536] 532. The method of aspect 531, wherein said factors known to be mitogenic for umbilical cord cells include one or more factors selected from a group comprising of: a) FGF-1; b) FGF-2; c) FGF-5; d) EGF; e) CNTF; f) KGF-1; g) PDGF; h) platelet rich plasma; i) TGF-alpha; and j) HGF-1.

[0537] 533. The method of aspect 528, wherein said umbilical cord cells are cultured under hypoxia.

[0538] 534. The method of aspect 528, wherein exosomes are collected from umbilical cord cells while said umbilical cord cells are in a proliferating state.

[0539] 535. The method of aspect 528, wherein said exosomes are collected from umbilical cord cells while said umbilical cord cells are cultured in a media containing no proliferative factors or largely reduced levels of said proliferation inducing growth factors.

[0540] 536. The method of aspect 528, wherein said exosomes are collected from said umbilical cord cells that have been cultured in 2-8% oxygen for at least 1 day.

[0541] 537. The method of aspect 536, wherein the cells are cultured for 1-15 days.

[0542] 538. The method of aspect 536, wherein the cells are cultured for 5-10 days.

[0543] 539. The method of aspect 536, wherein the cells are passaged for at least 1 passage.

[0544] 540. The method of aspect 536, wherein said exosomes are in a preparation, said preparation comprises less than 5% polyethylene glycol.

[0545] 541. The method of aspect 536, wherein the exosomes are purified using polyethylene glycol.

[0546] 542. The method of aspect 536, wherein the exosomes are purified using ultrafiltration.

[0547] 543. The method of aspect 536, wherein polyethylene glycol is added to the exosomes after purification.

[0548] 544. The method of aspect 528, wherein said exosomes express markers selected from a group comprising of: a) CD63; b) CD9; c) MHC I; d) CD56

[0549] 545. The method of aspect 528, wherein said umbilical cord cells are cultured in a media selected from a group comprising of: a) Roswell Park Memorial Institute (RPMI- 1640); b) Dublecco's Modified Essential Media (DMEM), c) Eagle's Modified Essential Media (EMEM), d) Optimem, and e) Iscove's Media.

[0550] 546. The method of aspect 528, wherein said exosomes are administered to a mammal in need of treatment suffering from a pulmonary disorder.

[0551] 547. The method of aspect 546, wherein said pulmonary disorder is associated with pulmonary inflammation.

[0552] 548. The method of aspect 547, wherein said pulmonary inflammation is associated with increased neutrophils in the lung area.

[0553] 549. The method of aspect 547, wherein said pulmonary inflammation is associated with increased mast cells in the lung area.

[0554] 550. The method of aspect 547, wherein said pulmonary inflammation is associated with increased T cells in the lung area.

[0555] 551. The method of aspect 547, wherein said pulmonary inflammation is associated with increased B cells in the lung area.

[0556] 552. The method of aspect 547, wherein said pulmonary inflammation is associated with increased NK cells in the lung area.

[0557] 553. The method of aspect 547, wherein said pulmonary inflammation is associated with increased gamma delta T cells in the lung area.

[0558] 554. The method of aspect 547, wherein said pulmonary inflammation is associated with increased monocytes in the lung area.

[0559] 555. The method of aspect 547, wherein said pulmonary inflammation is associated with increased complement activation in the lung area.

[0560] 556. The method of aspect 547, wherein said pulmonary inflammation is associated with increased coagulopathy in the lung area.

[0561] 557. The method of aspect 547, wherein an antifibrotic drug is administered together with said exosomes.

[0562] 558. The method of aspect 557 wherein said anti-fibrotic agent is relaxin-2, a recombinant form of relaxin-2 or a functional derivative or variant or mimetic of relaxin- 2.

[0563] 559. The method of aspect 558 wherein said recombinant form of relaxin-2 is serelaxin.

[0564] 560. The method of aspect 559 wherein the derivative of relaxin-2 is a single B chain derivative or an A and B chain truncate of relaxin-2.

[0565] 561. The method of aspect 560 wherein the single B chain derivative of relaxin-2 is H2-(B7-33) or the A and B chain truncate is H2-(A4-24)(B7-24).

[0566] 562. The method of aspect 557 further comprising the administration of anRXFP1 activating agent or agonist.

[0567]

[0568] 563. The method of aspect 557 wherein said umbilical cord derived cells are modified to express the relaxin or its derivative or variant.

[0569] 564. The method of aspect 557 wherein said umbilical cord exosomes are modified to contain the relaxin or its recombinant or derivative or variant form.

[0570] 565. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with relaxin.

[0571] 566. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with VEGF.

[0572] 567. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with EGF-1.

[0573] 568. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with IGF-1.

[0574] 569. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with PDGF-BB.

[0575] 570. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with hCG.

[0576] 571. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with FGF-1.

[0577] 572. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with FGF-2.

[0578] 573. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with FGF-5.

[0579] 574. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with TGF-beta.

[0580] 575. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with TGF-beta.

[0581] 576. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with interleukin 1 receptor antagonist.

[0582] 577. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with interleukin 4.

[0583] 578. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with angiopoietin.

[0584] 579. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with G-CSF.

[0585] 580. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with GM-CSF.

[0586] 581. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with M-CSF.

[0587] 582. The method of aspect 557 wherein said umbilical cord cells or exosomes are co-administered simultaneously or sequentially with low dose naltrexone.

[0588] 583. The method of aspects 557-582, wherein said umbilical cord cells, and / or exosomes and / or additives are administered intranasally, intramuscularly, intraperitoneally, intralymphatically, intraomentally, intrarespiratorily, intranasopharyngeally or intravenously administration.

[0589] 584. The method of aspect 557 wherein the airways disease is selected from the list consisting of asthma, allergic rhinitis, severe acute respiratory syndrome, COVID-19, virally associated lung damage, radiation associated lung damage, chronic obstructive pulmonary disease, pulmonary fibrosis, upper respiratory infection and reactive airways dysfunction syndrome.

[0590] 585. The method of aspect 557, wherein said umbilical cord cells, and / or exosomes are administered together with a cellular population capable of reducing inflammation.

[0591] 586. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a monocyte.

[0592] 587. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a T regulatory cell.

[0593] 588. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a Th2 cell.

[0594] 589. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a Th9 cell.

[0595] 590. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a Th3 cell.

[0596] 591. The method of aspect 585, wherein said cellular population capable of reducing inflammation is a Tri cell.

[0597] 592. The method of aspect 586, wherein said monocyte is a M2 monocyte.

[0598] 593. The method of aspect 592, wherein said M2 monocyte is derived from placenta, umbilical cord, bone marrow, wharton's jelly, or adipose derived.

[0599] 594. The method of aspect 593, wherein said M2 monocyte is cultured in the presence of interleukin 4.

[0600] 595. The method of aspect 593, wherein said M2 monocyte is cultured in the presence of interleukin 13.

[0601] 596. The method of aspect 593, wherein said M2 monocyte is cultured in the presence of interleukin 10.

[0602] 597. The method of aspect 593, wherein said M2 monocyte is capable of stimulating angiogenesis.

[0603] 598. The method of aspect 587, wherein said T regulatory cell expresses FOXP3.

[0604] 599. The method of aspect 587, wherein said T regulatory cell expresses membrane bound TGF-beta.

[0605] 600. The method of aspect 587, wherein said T regulatory cell is capable of suppressing dendritic cell maturation.

[0606] 601. The method of aspect 587, wherein said T regulatory cell is capable of suppressing proliferation of T cells.

[0607] 602. The method of aspect 587, wherein said T regulatory cell is capable of suppressing cytotoxicity of T cells.

[0608] 603. The method of aspect 587, wherein said T regulatory cell is capable of suppressing type 1 cytokine production of T cells.

[0609] 604. The method of aspect 603, wherein said type 1 cytokines are selected from a group comprising of: a) IL-2; b) IL-7; c) IL-12; d) IL-15; and e) IL-18.

[0610] 605. The method of aspect 588, wherein said Th2 cell expresses STAT-6.

[0611] 606. The method of aspect 588, wherein said Th2 cell expresses IL-4 or IL-13.

[0612] 607. The method of aspect 589, wherein said Th9 cell expresses IL-9.

[0613] 608. The method of aspect 590, wherein said Th3 cell expresses TGF-beta.

[0614] 609. The method of aspect 590, wherein said Th3 cell is associated with induction of oral tolerance.

[0615] 610. The method of aspect 590, wherein said Tri cells are associated with production of IL-10.

[0616] 611. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering from peripheral artery disease.

[0617] 612. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering from critical limb ischemia.

[0618] 613. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering from SARS-CoV-2 virus associated blood vessel dysfunction.

[0619] 614. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated with coagulopathy.

[0620] 615. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated with enhanced expression of tissue factor on endothelial cells.

[0621] 616. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated with enhanced endothelial cell activation.

[0622] 617. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated with reduced production of anti-thrombotic molecules on the endothelial surface.

[0623] 618. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated disseminated intravascular coagulation.

[0624] 619. The method of aspect 613, wherein said SARS-CoV-2 virus associated blood vessel dysfunction is associated with antiphospholipid antibody syndrome.

[0625] 620. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering from ischemic heart failure.

[0626] 621. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering post cardiac infarct scarring.

[0627] 622. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering post cerebral infarct scarring.

[0628] 623. The method of aspect 528, wherein said exosomes are used to stimulate new blood vessel formation in a patient suffering post cerebral infarct scarring.

[0629] 624. A method of stimulating regeneration in a tissue, wherein said in which regeneration is desired is distal from the tissue in which a regenerative means is administered through the steps of: a) identifying a diseased tissue in need of regeneration; and b) administering into said tissue a regenerative means in a manner so as to evoke a regenerative response in a tissue not treated with said regenerative means.

[0630] 625. The method of aspect 624, wherein said regeneration is desired in a tissue in which function has been lost or compromised.

[0631] 626. The method of aspect 624, wherein said tissue is comprised of one or more cells derived from the following: endothelial cells, epithelial cells, dermal cells, endodermal cells, mesodermal cells, fibroblasts, osteocytes, chondrocytes, natural killer cells, dendritic cells, hepatic cells, pancreatic cells, stromal cells, salivary gland mucous cells, salivary gland serous cells, von Ebner's gland cells, mammary gland cells, lacrimal gland cells, ceruminous gland cells, eccrine sweat gland dark cells, eccrine sweat gland clear cells, apocrine sweat gland cells, gland of Moll cells, sebaceous gland cells, bowman's gland cells, Brunner's gland cells, seminal vesicle cells, prostate gland cells, bulbourethral gland cells, Bartholin's gland cells, gland of Littre cells, uterus endometrium cells, isolated goblet cells, stomach lining mucous cells, gastric gland zymogenic cells, gastric gland oxyntic cells, pancreatic acinar cells, paneth cells, type IIpneumocytes, clara cells, somatotropes, lactotropes, thyrotropes, gonadotropes, corticotropes, intermediate pituitary cells, magnocellular neurosecretory cells, gut cells, respiratory tract cells, thyroid epithelial cells, parafollicular cells, parathyroid gland cells, parathyroid chief cell, oxyphil cell, adrenal gland cells, chromaffin cells, Leydig cells, theca interna cells, corpus luteum cells, granulosa lutein cells, theca lutein cells, juxtaglomerular cell, macula densa cells, peripolar cells, mesangial cell, blood vessel and lymphatic vascular endothelial fenestrated cells, blood vessel and lymphatic vascular endothelial continuous cells, blood vessel and lymphatic vascular endothelial splenic cells, synovial cells, serosal cell (lining peritoneal, pleural, and pericardial cavities), squamous cells, columnar cells, dark cells, vestibular membrane cell (lining endolymphatic space of ear), stria vascularis basal cells, stria vascularis marginal cell (lining endolymphatic space of ear), cells of Claudius, cells of Boettcher, choroid plexus cells, pia-arachnoid squamous cells, pigmented ciliary epithelium cells, nonpigmented ciliary epithelium cells, corneal endothelial cells, peg cells, respiratory tract ciliated cells, oviduct ciliated cell, uterine endometrial ciliated cells, rete testis ciliated cells, ductulus efferens ciliated cells, ciliated ependymal cells, epidermal keratinocytes, epidermal basal cells, keratinocyte of fingernails and toenails, nail bed basal cells, medullary hair shaft cells, cortical hair shaft cells, cuticular hair shaft cells, cuticular hair root sheath cells, hair root sheath cells of Huxley's layer, hair root sheath cells of Henle's layer, external hair root sheath cells, hair matrix cells, surface epithelial cells of stratified squamous epithelium, basal cell of epithelia, urinary epithelium cells, auditory inner hair cells of organ of Corti, auditory outer hair cells of organ of Corti, basal cells of olfactory epithelium, cold-sensitive primary sensory neurons, heat-sensitive primary sensory neurons, Merkel cells of epidermis, olfactory receptor neurons, pain-sensitive primary sensory neurons, photoreceptor rod cells, photoreceptor blue-sensitive cone cells, photoreceptor green-sensitive cone cells, photoreceptor red-sensitive cone cells, proprioceptive primary sensory neurons, touch-sensitive primary sensory neurons, type I carotid body cells, type II carotid body cell (blood pH sensor), type I hair cell of vestibular apparatus of ear (acceleration and gravity), type II hair cells of vestibular apparatus of ear, type I taste bud cells cholinergic neural cells, adrenergic neural cells, peptidergic neural cells, inner pillar cells of organ of Corti, outer pillar cells of organ of Corti, inner phalangeal cells of organ of Corti, outer phalangeal cells of organ of Corti, border cells of organ of Corti, Hensen cells of organ of Corti, vestibular apparatus supporting cells, taste bud supporting cells, olfactory epithelium supporting cells, Schwann cells, satellite cells,enteric glial cells, astrocytes, neurons, oligodendrocytes, spindle neurons, anterior lens epithelial cells, crystallin-containing lens fiber cells, hepatocytes, adipocytes, white fat cells, brown fat cells, liver lipocytes, kidney glomerulus parietal cells, kidney glomerulus podocytes, kidney proximal tubule brush border cells, loop of Henle thin segment cells, kidney distal tubule cells, kidney collecting duct cells, type I pneumocytes, pancreatic duct cells, nonstriated duct cells, duct cells, intestinal brush border cells, exocrine gland striated duct cells, gall bladder epithelial cells, ductulus efferens nonciliated cells, epididymal principal cells, epididymal basal cells, ameloblast epithelial cells, planum semilunatum epithelial cells, organ of Corti interdental epithelial cells, loose connective tissue fibroblasts, corneal keratocytes, tendon fibroblasts, bone marrow reticular tissue fibroblasts, nonepithelial fibroblasts, pericytes, nucleus pulposus cells, cementoblast / cementocytes, odontoblasts, odontocytes, hyaline cartilage chondrocytes, fibrocartilage chondrocytes, elastic cartilage chondrocytes, osteoblasts, osteocytes, osteoclasts, osteoprogenitor cells, hyalocytes, stellate cells (ear), hepatic stellate cells (Ito cells), pancreatic stelle cells, red skeletal muscle cells, white skeletal muscle cells, intermediate skeletal muscle cells, nuclear bag cells of muscle spindle, nuclear chain cells of muscle spindle, satellite cells, ordinary heart muscle cells, nodal heart muscle cells, Purkinje fiber cells, smooth muscle cells, myoepithelial cells of iris, myoepithelial cell of exocrine glands, reticulocytes, megakaryocytes, monocytes, connective tissue macrophages, epidermal Langerhans cells, dendritic cells, microglial cells, neutrophils, eosinophils, basophils, mast cell, helper T cells, suppressor ? cells, cytotoxic T cell, natural Killer T cells, B cells, natural killer cells, melanocytes, retinal pigmented epithelial cells, oogonia / oocytes, spermatids, spermatocytes, spermatogonium cells, spermatozoa, ovarian follicle cells, Sertoli cells, thymus epithelial cell, and / or interstitial kidney cells.

[0632] 627. The method of aspect 624, wherein said regenerative means is a growth factor or combination of growth factors.

[0633] 628. The method of aspect 627, wherein said growth factors are one or more selected from a group comprising of: AM, Ang, BMP, BDNF, EGF, Epo, FGF, GNDF, G-CSF, GM-CSF, GDF-9, HGF, HDGF, IGF, migration-stimulating factor, GDF-8, GDF-11, GDF-15, MGF, NGF, PIGF, PDGF, Tpo, TGF-.alpha., TGF-.beta., TNF-.alpha., VEGF, 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-12 35 kDa alpha subunit, IL-12 40 kDa beta subunit, IL-13, IL-14, IL-15, IL- 16, IL-17A, IL-17B, 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-28B, 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).

[0634] 629. The method of aspect 627, wherein said growth factor is platelet rich plasma.

[0635] 630. The method of aspect 629, wherein said platelet rich plasma is platelet lysate.

[0636] 631. The method of aspect 629, wherein said platelet rich plasma is derived from peripheral blood.

[0637] 632. The method of aspect 629, wherein said platelet rich plasma is derived from cord blood.

[0638] 633. The method of aspect 624, wherein said regenerative means comprises exosomes derived from a regenerative cell.

[0639] 634. The method of aspect 633, wherein said regenerative cell is a stem cell or a progenitor cell.

[0640] 635. The method of aspect 624, wherein said regenerative means is a mesenchymal stem cell.

[0641] 636. The method of aspect 635, wherein said mesenchymal stem cells are derived from a group of tissue sources selected from: a) foreskin; b) tummy tucks; c) placenta; d) ear lobe; e) adipose tissue; f) omentum; and g) wharton's jelly.

[0642] 637. The method of aspect 635, wherein said mesenchymal express markers selected from a group comprising of: a) NANOG; b) OCT-4; c) SSEA-4; and d) stem cell factor receptor.

[0643] 638. The method of aspect 624, wherein said regenerative means is a regenerative cell.

[0644] 639. The method of 638, wherein said stem cells are pluripotent stem cells.

[0645] 640. The method of aspect 639, wherein said pluripotent stem cells are selected from a group comprising of: a) embryonic stem cells; b) parthenogenic derived stem cells; c) inducible pluripotent stem cells; d) somatic cell nuclear transfer derived stem cells; e) cytoplasmic transfer derived stem cells; and f) stimulus-triggered acquisition of pluripotency.

[0646] 641. The method of 634, wherein said stem cells are hematopoietic stem cell.

[0647] 642. The method of aspect 641, wherein said hematopoietic stem cells are capable of multi-lineage reconstitution in an immunodeficient host.

[0648] 643. The method of aspect 641, wherein said hematopoietic stem cells express the c-kit protein.

[0649] 644. The method of aspect 641, wherein said hematopoietic stem cells express the Sca-1 protein.

[0650] 645. The method of aspect 641, wherein said hematopoietic stem cells expressCD34.

[0651] 646. The method of aspect 641, wherein said hematopoietic stem cells expressCD133.

[0652] 647. The method of aspect 641, wherein said hematopoietic stem cells lack expression of lineage markers.

[0653] 648. The method of aspect 641, wherein said hematopoietic stem cells lack expression of CD38.

[0654] 649. The method of aspect 641, wherein said hematopoietic stem cells are positive for expression of c-kit and Sca-1 and substantially lack expression of lineage markers.

[0655] 650. The method of aspect 641, wherein said hematopoietic stem cells are derived from a group of sources, said group comprising of: a) peripheral blood; b) mobilized peripheral blood; c) bone marrow; d) cord blood; e) adipose stromal vascular fraction; and f) derived from progenitor cells.

[0656] 651. The method of aspect 643, wherein said progenitor cell is a pluripotent stem cell.

[0657] 652. The method of aspect 641, wherein said stem cells are mesenchymal stem cells.

[0658] 653. The method of aspect 652, wherein said mesenchymal stem cells are plastic adherent.

[0659] 654. The method of aspect 652, wherein said mesenchymal stem cells express a marker selected from a group comprising of: a) CD73; b) CD90; and c) CD105.

[0660] 655. The method of aspect 652, wherein said mesenchymal stem cells lack expression of a marker selected from a group comprising of: a) CD14; b) CD45; and c) CD34.

[0661] 656. The method of aspect 652, wherein said mesenchymal stem cells are derived from tissues selected from a group comprising of: a) bone marrow; b) peripheral blood; c) adipose tissue; d) mobilized peripheral blood; e) umbilical cord blood; f) Wharton's jelly; g) umbilical cord tissue; h) skeletal muscle tissue; i) subepithelial umbilical cord; j) endometrial tissue; k) menstrual blood; and I) fallopian tube tissue.

[0662] 657. The method of aspect 656, wherein said mesenchymal stem cells from umbilical cord tissue express markers selected from a group comprising of; a) oxidized low density lipoprotein receptor 1, b) chemokine receptor ligand 3; and c) granulocyte chemotactic protein.

[0663] 658. The method of aspect 656, wherein said mesenchymal stem cells from umbilical cord tissue do not express markers selected from a group comprising of: a) CD117; b) CD31; c) CD34; and CD45;

[0664] 659. The method of aspect 656, wherein said mesenchymal stem cells from umbilical cord tissue express, relative to a human fibroblast, increased levels of interleukin 8 and reticulon 1

[0665] 660. The method of aspect 656, wherein said mesenchymal stem cells from umbilical cord tissue have the potential to differentiate into cells of at least a skeletal muscle, vascular smooth muscle, pericyte or vascular endothelium phenotype.

[0666] 661. The method of aspect 656, wherein said mesenchymal stem cells from umbilical cord tissue express markers selected from a group comprising of: a) CD10; b) CD13; c) CD44; d) CD73; and e) CD90.

[0667] 662. The method of aspect 656, wherein said umbilical cord tissue mesenchymal stem cell is an isolated umbilical cord tissue cell isolated from umbilical cord tissue substantially free of blood that is capable of self-renewal and expansion in culture,

[0668] 663. The method of aspect 656, wherein said umbilical cord tissue mesenchymal stem cells has the potential to differentiate into cells of other phenotypes.

[0669] 664. The method of aspect 656, wherein said other phenotypes comprise: a) osteocytic; b) adipogenic; and c) chondrogenic differentiation.

[0670] 665. The method of aspect 656, wherein said cord tissue derived mesenchymal stem cells can undergo at least 20 doublings in culture.

[0671] 665. The method of aspect 656, wherein said cord tissue derived mesenchymal stem cell maintains a normal karyotype upon passaging

[0672] 667. The method of aspect 656, wherein said cord tissue derived mesenchymal stem cell expresses a marker selected from a group of markers comprised of: a) CD10 b) CD13; c) CD44; d) CD73; e) CD90; f) PDGFr-alpha; g) PD-L2; and h) HLA-A,B,C

[0673] 668. The method of aspect 656, wherein said cord tissue mesenchymal stem cells does not express one or more markers selected from a group comprising of; a) CD31; b) CD34; c) CD45; d) CD80; e) CD86; f) CD117; g) CD141; h) CD178; i) B7-H2; j) HLA-G and k) H LA-DR, DP, DQ.

[0674] 669. The method of aspect 656, wherein said umbilical cord tissue-derived cell secretes factors selected from a group comprising of: a) MCP-l; b) MIPIbeta; c) IL-6; d) IL-8; e) GCP-2; f) HGF; g) KGF; h) FGF; i) HB-EGF; j) BDNF; k) TPO; I) RANTES; and m) TIMP1

[0675] 670. The method of aspect 656, wherein said umbilical cord tissue derived cells express markers selected from a group comprising of: a) TRA1-60; b) TRA1-81; c) SSEA3; d) SSEA4; and e) NANOG.

[0676] 671. The method of aspect 656, wherein said umbilical cord tissue-derived cells are positive for alkaline phosphatase staining.

[0677] 672. The method of aspect 656, wherein said umbilical cord tissue-derived cells are capable of differentiating into one or more lineages selected from a group comprising of; a) ectoderm; b) mesoderm, and; c) endoderm.

[0678] 673. The method of aspect 656, wherein said bone marrow derived mesenchymal stem cells possess markers selected from a group comprising of: a) CD73; b) CD90; and c) CD105.

[0679] 674. The method of aspect 656, wherein said bone marrow derived mesenchymal stem cells possess markers selected from a group comprising of: a) LFA-3; b) ICAM-1; c) PECAM-1; d) P-selectin; e) L-selectin; f) CD49b / CD29; g) CD49c / CD29; h) CD49d / CD29; i) CD29; j) CD18; k) CD61; I) 6-19; m) thrombomodulin; n) telomerase; o) CD10; p) CD13; and q) integrin beta.

[0680] 675. The method of aspect 656, wherein said bone marrow derived mesenchymal stem cell is a mesenchymal stem cell progenitor cell.

[0681] 676. The method of aspect 675, wherein said mesenchymal progenitor cells are a population of bone marrow mesenchymal stem cells enriched for cells containing STRO-1

[0682] 677. The method of aspect 676, wherein said mesenchymal progenitor cells express both STRO-1 and VCAM-l.

[0683] 678. A method of aspect 676, wherein said STRO-1 expressing cells are negative for at least one marker selected from the group consisting of: a) CBFA-1; b) collagen type II; c) PPAR.gamma2; d) osteopontin; e) osteocalcin; f) parathyroid hormone receptor; g) leptin; h) H-ALBP; i) aggrecan; j) Ki67, and k) glycophorin A.

[0684] 679. The method of aspect 675, wherein said bone marrow mesenchymal stem cells lack expression of CD14, CD34, and CD45.

[0685] 680. The method of aspect 678, wherein said STRO-1 expressing cells are positive for a marker selected from a group comprising of: a) VACM-l; b) TKY-1; c) CD146 and; d) STRO-2

[0686] 681. The method of aspect 656, wherein said bone marrow mesenchymal stem cell express markers selected from a group comprising of: a) CD13; b) CD34; c) CD56 and; d) CD117

[0687] 682. The method of aspect 656, wherein said bone marrow mesenchymal stem cells do not express CD10.

[0688] 683. The method of aspect 656, wherein said bone marrow mesenchymal stem cells do not express CD2, CD5, CD14, CD19, CD33, CD45, and DRII.

[0689] 684. The method of aspect 656, wherein said bone marrow mesenchymal stem cells express CD13,CD34, CD56, CD90, CD117 and nestin, and which do not express CD2, CD3, CD10, CD14, CD16, CD31, CD33, CD45 and CD64.

[0690] 685. The method of aspect 656, wherein said skeletal muscle stem cells express markers selected from a group comprising of: a) CD13; b) CD34; c) CD56 and; d) CD117

[0691] 686. The method of aspect 685, wherein said skeletal muscle mesenchymal stem cells do not express CD10.

[0692] 687. The method of aspect 685, wherein said skeletal muscle mesenchymal stem cells do not express CD2, CD5, CD14, CD19, CD33, CD45, and DRII.

[0693] 688. The method of aspect 656, wherein said subepithelial umbilical cord derived mesenchymal stem cells possess markers selected from a group comprising of; a) CD29; b) CD73; c) CD90; d) CD166; e) SSEA4; f) CD9; g) CD44; h) CD146; and i) CD105

[0694] 689. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells do not express markers selected from a group comprising of; a)CD45; b) CD34; c) CD14; d) CD79; e) CD106; f) CD86; g) CD80; h) CD19; i) CD117; j) Stro-1 and k) HLA-DR.

[0695] 690. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells express CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, and CD105.

[0696] 691. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells do not express CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, and HLA-DR.

[0697]

[0698] 692. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells are positive for SOX2.

[0699] 693. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells are positive for OCT4.

[0700] 694. The method of aspect 688, wherein said subepithelial umbilical cord derived mesenchymal stem cells are positive for OCT4 and SOX2.

[0701] 695. The method of aspect 624, wherein said regenerative means comprises umbilical cord cell derived apoptotic vesicles.

[0702] 696. The method of aspect 624, wherein said regenerative means comprises umbilical cord cell derived miRNAs.

[0703] 697. The method of aspect 633, wherein said exosomes possess a size of between 30 nm and 150 nm.

[0704] 698. The method of aspect 697, wherein said exosome possesses a size of between 2 nm and 200 nm, as determined by filtration against a 0.2 .mu.M filter and concentration against a membrane with a molecular weight cut-off of 10 kDa, or a hydrodynamic radius of below 100 nm as determined by laser diffraction or dynamic light scattering.

[0705] 699. The method of aspect 633, wherein said exosome possesses a lipid selected from the group consisting of: a) phospholipids; b) phosphatidyl serine; c) phosphatidyl inositol; d) phosphatidyl choline; e) sphingomyelin; f) ceramides; g) glycolipid; h) cerebroside; i) steroids, and j) cholesterol.

[0706] 700. The method of aspect 633, wherein said exosome possesses a lipid raft.

[0707] 701. The method of aspect 633, wherein said exosome expresses antigenic markers on surface of said exosome, wherein said antigenic markers are selected from a group comprising of: a) CD9; b) CD63; c) CD81; d) ANXA2; e) ENO1; f) HSP90AA1; g) EEF1A1; h) YWHAE; i) SDCBP; j) PDCD6IP; k) ALB; I) YWHAZ; m) EEF2; n) ACTG1; o) LDHA; p) HSP90AB1; q) ALDOA; r) MSN; s) ANXA5; t) PGK1; and u) CFL1.

[0708] 702. The method of aspect 624, wherein enhancement of distant regenerative effect is accomplished by systemic administration of an epigenetic acting drug.

[0709] 703. The method of aspect 702, wherein said epigenetic acting drug is a histone deacetylase inhibitor.

[0710] 704. The method of aspect 702, wherein said epigenetic acting drug is a DNA methyltransferase inhibitor.

[0711] 705. The method of aspect 633, wherein said exosomes are utilized to reprogram another cell type in vivo, in vitro, or ex vivo, wherein reprogramming by said exosomes endows said cell type with a therapeutic property.

[0712] 706. The method of aspect 705, wherein said exosomes are incubated with T cells in order to generate T cells with therapeutic properties.

[0713] 707. The method of aspect 706, wherein said T cells are endowed with ability to stimulate angiogenesis after culture with said umbilical cord derived exosomes.

[0714] 708. The method of aspect 706, wherein said T cells are endowed with ability to stimulate neurogenesis after culture with said umbilical cord derived exosomes.

[0715] 709. The method of aspect 706, wherein said T cells are endowed with ability to stimulate suppression of inflammation after culture with said umbilical cord derived exosomes.

[0716] 710. The method of aspect 706, wherein said T cells are endowed with ability to stimulate neutrophil apoptosis after culture with said umbilical cord derived exosomes.

[0717] 711. The method of aspect 706, wherein said T cells are endowed with ability to suppress maturation of dendritic cells after culture with said umbilical cord derived exosomes.

[0718] 712. The method of aspect 706, wherein said T cells are endowed with ability to suppress generation of interleukin-17 after culture with said umbilical cord derived exosomes.

[0719] 713. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage angiogenic activity.

[0720] 714. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage antifibrotic activity.

[0721] 715. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage neurogenic activity.

[0722] 716. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage anti-apoptotic activity.

[0723] 717. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage wound healing activity.

[0724] 718. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage anti-inflammatory activity.

[0725] 719. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage ability to induce neutrophil apoptosis.

[0726] 720. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage ability to suppress dendritic cell maturation.

[0727] 721. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage ability to suppress generation of Thl cells.

[0728] 722. The method of aspect 721, wherein said Thl cells express Helios.

[0729] 723. The method of aspect 721, wherein said Thl cells express interferon gamma.

[0730] 724. The method of aspect 721, wherein said Thl cells express STAT4.

[0731] 725. The method of aspect 705, wherein said exosomes are utilized to reprogram a macrophage to endow onto said macrophage ability to stimulate generation of T regulatory cells.

[0732] 726. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting inflammation.

[0733]

[0734] 727. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting autoimmunity.

[0735] 728. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting multiple organ failure.

[0736] 729. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting acute respiratory distress syndrome.

[0737] 730. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting liver failure.

[0738] 731. The method of aspect 725, wherein said T regulatory cells are capable of inhibiting pulmonary fibrosis.

[0739] 732. The method of aspect 705, wherein said exosomes are used to reprogram a macrophage, wherein said reprogrammed macrophage can be used for treatment of an orthopedic injury.

[0740] 733. The method of aspect 732, wherein said reprogrammed macrophage is aM2 macrophage.

[0741] 734. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of VEGF as compared on a macrophage that has not been reprogrammed.

[0742] 735. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of FGF-1 as compared on a macrophage that has not been reprogrammed.

[0743] 736. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of FGF-2 as compared on a macrophage that has not been reprogrammed.

[0744] 737. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of FGF-5 as compared on a macrophage that has not been reprogrammed.

[0745] 738. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of HGF-1 as compared on a macrophage that has not been reprogrammed.

[0746] 739. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of PDGF-1 as compared on a macrophage that has not been reprogrammed.

[0747] 740. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of interleukin 1 receptor antagonist as compared on a macrophage that has not been reprogrammed.

[0748] 741. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of interleukin 1 receptor antagonist as compared on a macrophage that has not been reprogrammed.

[0749] 742. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of interleukin-10 as compared on a macrophage that has not been reprogrammed.

[0750] 743. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of PGE-2 as compared on a macrophage that has not been reprogrammed.

[0751] 744. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of indolamine 2,3 deoxygenase as compared on a macrophage that has not been reprogrammed.

[0752] 745. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of Galectin 3 as compared on a macrophage that has not been reprogrammed.

[0753] 746. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of Galectin 9 as compared on a macrophage that has not been reprogrammed.

[0754] 747. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of HLA-G as compared on a macrophage that has not been reprogrammed.

[0755] 748. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of I LT-3 as compared on a macrophage that has not been reprogrammed.

[0756] 749. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of TIGIT as compared on a macrophage that has not been reprogrammed.

[0757] 750. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of arginase as compared on a macrophage that has not been reprogrammed.

[0758] 751. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of IL-6 as compared on a macrophage that has not been reprogrammed.

[0759] 752. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of serpin-1 as compared on a macrophage that has not been reprogrammed.

[0760] 753. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of PD-L1 as compared on a macrophage that has not been reprogrammed.

[0761] 754. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of PD-L2 as compared on a macrophage that has not been reprogrammed.

[0762] 755. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially higher amount of CD206 as compared on a macrophage that has not been reprogrammed.

[0763] 756. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially lower amount of eotoxin as compared on a macrophage that has not been reprogrammed.

[0764] 757. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially lower amount of CD14 as compared on a macrophage that has not been reprogrammed.

[0765] 758. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially lower amount of M-CSF receptor as compared on a macrophage that has not been reprogrammed.

[0766] 759. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially lower amount of TGF-alpha as compared on a macrophage that has not been reprogrammed.

[0767] 760. The method of aspect 732, wherein said reprogrammed macrophage expresses a substantially lower amount of interleukin 13 as compared on a macrophage that has not been reprogrammed.

[0768] 761. The method of aspect 732, wherein said population of reprogrammed macrophage cells is administered by injection.

[0769] 762. The method of aspect 761, wherein the population of cells is administered by injection with a pharmaceutically acceptable carrier.

[0770] 763. The method of aspect 762, wherein said pharmaceutically acceptable carrier maintains viability and function of said reprogrammed macrophages.

[0771] 764. The method of aspect 762, wherein said pharmaceutically acceptable carrier is hyaluronic acid.

[0772] 765. The method of aspect 762, wherein said pharmaceutically acceptable carrier is decellularized tissue.

[0773] 766. The method of aspect 765, wherein said decellularized tissue is placental tissue.

[0774] 767. The method of aspect 765, wherein said decellularized tissue is omentum tissue.

[0775] 768. The method of aspect 765, wherein said decellularized tissue is adipose tissue.

[0776] 769. The method of aspect 765, wherein said decellularized tissue is bone marrow tissue.

[0777] 770. The method of aspect 765, wherein said decellularized tissue is subintestinal mucosa tissue.

[0778] 771. The method of aspect 762, wherein said cells are administered together with an antioxidant.

[0779] 772. The method of aspect 762, wherein said cells are administered together with a growth factor.

[0780] 773. The method of aspect 772, wherein said growth factor is TGF-beta.

[0781] 774. The method of aspect 772, wherein said growth factor is VEGF.

[0782] 775. The method of aspect 772, wherein said growth factor is EGF.

[0783] 776. The method of aspect 772, wherein said growth factor is NGF.

[0784] 777. The method of aspect 772, wherein said growth factor is HGF-1.

[0785] 778. The method of aspect 772, wherein said growth factor is bone morphogenic protein-1.

[0786] 779. The method of aspect 772, wherein said growth factor is bone morphogenic protein-2.

[0787] 780. The method of aspect 772, wherein said growth factor is platelet rich plasma.

[0788] 781. The method of aspect 772, wherein said growth factor is conditioned media from a regenerative cell treated with an inflammatory stimuli.

[0789] 782. The method of aspect 781, wherein said regenerative cell is a mesenchymal stem cell.

[0790] 783. The method of aspect 781, wherein said regenerative cell is cord blood mononuclear cells.

[0791] 784. The method of aspect 781, wherein said regenerative cell is peripheral blood mononuclear cells.

[0792] 785. The method of aspect 781, wherein said regenerative cell is bone marrow mononuclear cells.

[0793] 786. The method of aspect 781, wherein said regenerative cell is stromal vascular fraction cells.

[0794] 787. The method of aspect 781, wherein said regenerative cell is a mixed lymphocyte reaction.

[0795] 788. The method of aspect 781, wherein said regenerative cell is thymic medullary epithelial cells.

[0796] 789. The method of aspect 781, wherein said regenerative cell is dendritic cells.

[0797] 790. The method of aspect 781, wherein said inflammatory stimuli is a toll like receptor agonist.

[0798] 791. The method of aspect 790, wherein said toll like receptor is TLR-1.

[0799] 792. The method of aspect 791, wherein said activator of TLR-1 is Pam3CSK4.

[0800] 793. The method of aspect 790, wherein said toll like receptor is TLR-2.

[0801] 794. The method of aspect 793, wherein said activator of TLR-2 is HKLM.

[0802] 795. The method of aspect 790, wherein said toll like receptor is TLR-3.

[0803] 796. The method of aspect 795, wherein said activator of TLR-3 is Poly:IC.

[0804] 797. The method of aspect 790, wherein said toll like receptor is TLR-4.

[0805] 798. The method of aspect 797, wherein said activator of TLR-4 is LPS.

[0806] 799. The method of aspect 797, wherein said activator of TLR-4 isBuprenorphine.

[0807] 800. The method of aspect 797, wherein said activator of TLR-4 isCarbamazepine.

[0808] 801. The method of aspect 797wherein said activator of TLR-4 is Fentanyl.

[0809] 802. The method of aspect 797, wherein said activator of TLR-4 is Levorphanol.

[0810] 803. The method of aspect 797, wherein said activator of TLR-4 is Methadone.

[0811] 804. The method of aspect 797, wherein said activator of TLR-4 is Cocaine.

[0812] 805. The method of aspect 797, wherein said activator of TLR-4 is Morphine.

[0813] 806. The method of aspect 797, wherein said activator of TLR-4 isOxcarbazepine.

[0814] 807. The method of aspect 797, wherein said activator of TLR-4 is Oxycodone.

[0815] 808. The method of aspect 797, wherein said activator of TLR-4 is Pethidine.

[0816] 809. The method of aspect 797, wherein said activator of TLR-4 isGlucuronoxylomannan from Cryptococcus.

[0817] 810. The method of aspect 797, wherein said activator of TLR-4 is Morphine-3- glucuronide.

[0818] 811. The method of aspect 797, wherein said activator of TLR-4 is lipoteichoic acid.

[0819] 812. The method of aspect 797, wherein said activator of TLR-4 is -defensin 2.

[0820] 813. The method of aspect 797, wherein said activator of TLR-4 is small molecular weight hyaluronic acid.

[0821] 814. The method of aspect 797, wherein said activator of TLR-4 is fibronectinEDA.

[0822] 815. The method of aspect 797, wherein said activator of TLR-4 is snapin.

[0823] 816. The method of aspect 797 , wherein said activator of TLR-4 is tenascin C.

[0824] 817. The method of aspect 790, wherein said toll like receptor is TLR-5.

[0825] 818. The method of aspect 817, wherein said activator of TLR-5 is f lagell in.

[0826] 819. The method of aspect 790, wherein said toll like receptor is TLR-6.

[0827] 820. The method of aspect 819, wherein said activator of TLR-6 is FSL-1.

[0828] 821. The method of aspect 790, wherein said toll like receptor is TLR-7.

[0829] 822. The method of aspect 821, wherein said activator of TLR-7 is imiquimod.

[0830] 823. The method of aspect 790, wherein said toll like receptor of TLR-8.

[0831] 825. The method of aspect 823, wherein said activator of TLR8 is ssRNA40 / LyoVec.

[0832] 826. The method of aspect 790, wherein said toll like receptor of TLR-9.

[0833] 827. The method of aspect 826, wherein said activator of TLR-9 is a CpG oligonucleotide.

[0834] 828. The method of aspect 827, wherein said activator of TLR-9 is ODN2006.

[0835] 829. The method of aspect 827, wherein said activator of TLR-9 is Agatolimod.

[0836] 830. The method of aspect 1, wherein the population of cells is administered surgically.

[0837] 831. The method of aspect 732, wherein said orthopedic injury is selected from the group consisting of a partial tendon tear, a complete tendon tear, a partial tendon laceration, a compete tendon laceration, a partial tendon avulsion, a complete tendon avulsion, a partial ligament tear, a complete ligament tear, a partial ligament laceration, a compete ligament laceration, tendinopathy, tendinosis, tendinitis, meniscal tears, joint capsule tears.

[0838] 832. The method of aspect 732, wherein the orthopedic injury is selected from the group consisting of plantar fasciitis, tennis elbow, bicep tendinitis, and carpal tunnel syndrome.

[0839] 833. The method of aspect 732, wherein the population of reprogrammed macrophages is generated by a method comprising the step of: co-culturing a CD56 expressing umbilical cord mesenchymal stem cell with a monocyte in a manner in which said monocyte acquires anti-inflammatory properties.

[0840] 834. The method of aspect 833, wherein said culture is performed in the presence of hypoxia.

[0841] 835. The method of aspect 834, wherein said culture is performed in the presence of valproic acid.

[0842] 836. The method of aspect 834, wherein said culture is performed in the presence of valproic acid.

[0843] 837. The method of aspect 834, wherein said culture is performed in the presence of lithium.

[0844] 838. The method of aspect 834, wherein said culture is performed in the presence of GM-CSF.

[0845] 839. The method of aspect 834, wherein said culture is performed in the presence of CNTF.

[0846] 840. The method of aspect 834, wherein said culture is performed in the presence of trichostatin-A.

[0847] 841. The method of aspect 834, wherein said culture is performed in the presence of an antioxidant.

[0848] 842. The method of aspect 841, wherein said antioxidant is selected from a group comprising of: a) ascorbic acid; b) n-acetylcysteine; c) glutathione; d) superoxide dismutase; e) rutin; f) pterostilbene; g) resveratrol; h) vitamin A; i) vitamin D; j) vitamin E; k) fish oil; and I) quercetin.

[0849] 843. The method of aspect 834, wherein said culture is performed in the presence of an inhibitor of NF-kappa B.

[0850] 844. The method of aspect 843, wherein said inhibitor of NF-kappa B is selected from a group comprising of: Calagualine (fern derivative), Conophylline (Ervatamia microphylla), Evodiamine (Evodiae fructus component), Geldanamycin, Perrilyl alcohol, Protein-bound polysaccharide from basidiomycetes, Rocaglamides (Aglaia derivatives), 15-deoxy-prostaglandin J(2), Lead, Anandamide, Artemisia vestita, Cobrotoxin, Dehydroascorbic acid (Vitamin C), Herbimycin A, Isorhapontigenin, Manumycin A, Pomegranate fruit extract, Tetrandine (plant alkaloid), Thienopyridine, Acetyl-boswellic acids, l'-Acetoxychavicol acetate (Languas galanga), Apigenin (plant flavinoid),Cardamomin, Diosgenin, Furonaphthoquinone, Guggulsterone, Falcarindol, Honokiol, Hypoestoxide, Garcinone B, Kahweol, Kava (Piper methysticum) derivatives, mangostin (from Garcinia mangostana), N-acetylcysteine, Nitrosylcobalamin (vitamin B12 analog), Piceatannol, Plumbagin (5-hydroxy-2-methyl-l,4-naphthoquinone), Quercetin, Rosmarinic acid, Semecarpus anacardiu extract, Staurosporine, Sulforaphane and phenylisothiocyanate, Theaflavin (black tea component), Tilianin, Tocotrienol, Wedelolactone, Withanolides, Zerumbone, Silibinin, Betulinic acid, Ursolic acid, Monochloramine and glycine chloramine (NH2CI), Anethole, Baoganning, Black raspberry extracts (cyanidin 3-O-glucoside, cyanidin 3-O-(2(G)-xylosylrutinoside), cyanidin 3-O-rutinoside), Buddlejasaponin IV, Cacospongionolide B, Calagualine, Carbon monoxide, Cardamonin, Cycloepoxydon; l-hydroxy-2-hydroxymethyl-3-pent-l- enylbenzene, Decursin, Dexanabinol, Digitoxin, Diterpenes, Docosahexaenoic acid, Extensively oxidized low density lipoprotein (ox-LDL), 4-Hydroxynonenal (HNE), Flavopiridol, [6]-gingerol; casparol, Glossogyne tenuifolia, Phytic acid (inositol hexakisphosphate), Pomegranate fruit extract, Prostaglandin Al, 20(S)-Protopanaxatriol (ginsenoside metabolite), Rengyolone, Rottierin, Saikosaponin-d, Saline (low Na+ istonic)

[0851] 845. The method of aspect 834, wherein said culture is performed in the presence of low level laser irradiation.

[0852] 846. The method of aspect 845, wherein said low level laser irradiation is provided in at least one wavelength, said wavelength in a range between about 620 nanometers and about 1070 nanometers.

[0853] 847. The method of aspect 845, wherein said laser irradiation is administered by a light source between approximately 100 .mu.W / cm.sup.2 to approximately 10 W / cm.sup.2.

[0854] 848. The method of aspect 845, wherein said low level laser irradiation enhances growth factor production of said cells.

[0855] 849. The method of aspect 845, wherein said low level laser irradiation enhances chemotactic activityof said cells.

[0856] 850. The method of aspect 849, wherein said chemotactic ability is correlated with expression of the chemokine receptor CXCR-4, which is a membrane bound receptor whose ligand is SDF-1, otherwise known as CXCL12.

[0857] 851. A method of generating therapeutic macrophages comprising the steps of: a) obtaining a monocyte or monocyte progenitor cell; and b) contacting said monocyte or monocytic progenitor cell with a pluripotent stem cell for a sufficient time period toendow said monocyte or monocytic progenitor cell with ability to stimulate angiogenesis.

[0858] 852. The method of aspect 851, wherein said monocyte cells are derived from peripheral blood mononuclear cells.

[0859] 853. The method of aspect 852, wherein said monocyte cells express CD14.

[0860] 854. The method of aspect 852, wherein said monocyte cells express CD16.

[0861] 855. The method of aspect 852, wherein said monocyte cells are plastic adherent.

[0862] 856. The method of aspect 852, wherein said monocyte cells express CDllb.

[0863] 857. The method of aspect 852, wherein said monocytic progenitor is a myeloid progenitor cell.

[0864] 858. The method of aspect 851, wherein said monocyte cells are derived from bone marrow.

[0865] 859. The method of aspect 858, wherein said monocyte cells express CD14.

[0866] 860. The method of aspect 858, wherein said monocyte cells express CD16.

[0867] 861. The method of aspect 858, wherein said monocyte cells are plastic adherent.

[0868] 862. The method of aspect 858, wherein said monocyte cells express CDllb.

[0869] 863. The method of aspect 858, wherein said monocytic progenitor is a myeloid progenitor cell.

[0870] 864. The method of aspect 851, wherein said monocytes are derived from mobilized peripheral blood.

[0871] 865. The method of aspect 864, wherein said mobilization of peripheral blood is accomplished through pretreatment of the patient with G-CSF.

[0872] 866. The method of aspect 864, wherein said mobilization of peripheral blood is accomplished through pretreatment of the patient with flt-3 ligand.

[0873] 867. The method of aspect 864, wherein said mobilization of peripheral blood is accomplished through pretreatment of the patient with Mozibil.

[0874] 868. The method of aspect 864, wherein said mobilization of peripheral blood is accomplished through pretreatment of the patient with Mozibil.

[0875] 869. The method of aspect 864, wherein said monocyte cells express CD14.

[0876] 870. The method of aspect 864, wherein said monocyte cells express CD16.

[0877] 871. The method of aspect 864, wherein said monocyte cells are plastic adherent.

[0878] 872. The method of aspect 864, wherein said monocyte cells express CDllb.

[0879] 873. The method of aspect 851, wherein said pluripotent stem cell is derived from Wharton's Jelly and is derived from either the patient to be treated (autologous) or said donor is different from the patient to be treated (allogeneic).

[0880] 874. The method of aspect 873, wherein said pluripotent stem cells are cultured in a media allowing for stem cell proliferation.

[0881] 875. The method of aspect 874, wherein said media allowing for pluripotent stem cell proliferation contains one or more factors known to be mitogenic for mesenchymal stem cells.

[0882] 876. The method of aspect 875, wherein said factors known to be mitogenic for mesenchymal stem cells include one or more factors selected from a group comprising of: a) FGF-1; b) FGF-2; c) FGF-5; d) EGF; e) CNTF; f) KGF-1; g) PDGF; h) platelet rich plasma; i) TGF-alpha; and j) HGF-1.

[0883] 877. The method of aspect 873, wherein said stem cells are cultured under hypoxia.

[0884] 878. The method of aspect 851, wherein said stem cells are exposed to allogeneic T cells to induce enhancement of regenerative activity.

[0885] 879. The method of aspect 851, wherein said stem cells are exposed to extracorporeal pulse wave ultrasound in order to induce expression of stress related proteins such as hsp90. .

[0886] 880. The method of aspect 851, wherein said stem cells are treated with an inflammatory stimuli to mimic a wound environment.

[0887] 881. The method of aspect 880, wherein said inflammatory stimuli is an inflammatory cytokine.

[0888] 882. The method of aspect 881, wherein said inflammatory cytokine is TNF- alpha.

[0889] 883. The method of aspect 882, wherein said inflammatory cytokine is IL-1.

[0890] 884. The method of aspect 883, wherein said inflammatory cytokine is IL-6.

[0891] 885. The method of aspect 883, wherein said inflammatory cytokine is IL-11.

[0892] 886. The method of aspect 883, wherein said inflammatory cytokine is IL-12. .

[0893] 887. The method of aspect 883, wherein said inflammatory cytokine is IL-17. .

[0894] 888. The method of aspect 883, wherein said inflammatory cytokine is IL-18. .

[0895] 889. The method of aspect 883, wherein said inflammatory cytokine is IL-21. .

[0896] 890. The method of aspect 883, wherein said inflammatory cytokine is IL-33. .

[0897] 891. The method of aspect 881, wherein said inflammatory cytokine is capable of stimulating expression of genes in fibroblast cells selected from a group comprising of: IL-6, Myosin 1, IL-33, Hypoxia Inducible Factor-1, Guanylate Binding Protein Isoform I, Aminolevulinate delta synthase 2, AMP deaminase, IL-17, DNAJ-like 2 protein, Cathepsin L, Transcription factor-20, M31724, pyenylalkylamine binding protein; HEC, GA17, arylsulfatase D gene, arylaulfatase E gene, cyclin protein gene, pro-platelet basic protein gene, PDGFRA, human STS WI-12000, mannosidase, beta A, lysosomal MAN BA gene, UBE2D3 gene, Human DNA for Ig gamma heavy-chain, STRL22, BHMT, homo sapiens Down syndrome critical region, FI5613 containing ZNF gene family member, IL8, ELFR, homo sapiens mRNA for dual specificity phosphatase MKP-5, homo sapiens regulator of G protein signaling 10 mRNA complete, Homo sapiens Wnt-13 Mma, homo sapiens N- terminal acetyltransferase complex ardl subunit, ribosomal protein L15 mRNA, PCNA mRNA, ATRM gene exon 21, HR gene for hairless protein exon 2, N-terminal acetyltransferase complex ard 1 subunit, HSM801431 homo sapiens mRNA, CDNA DKFZp434N2072,RPL26, and HR gene for hairless protein, regulator of G protein signaling

[0898] 892. The method of aspect 851, wherein said monocytes or monocytic progenitors are cultured with said stem cells at a ratio of 1 monocyte or monocytic progenitor cell to 100 stem cells.

[0899] 893. The method of aspect 892, wherein said monocytes or monocytic progenitors are cultured with said stem cells at a ratio of 1 monocyte or monocytic progenitor cell to 10 stem cells.

[0900] 894. The method of aspect 893, wherein said monocytes or monocytic progenitors are cultured with said stem cells at a ratio of 1 monocyte or monocytic progenitor cell to 10 stem cells.

[0901] 895. The method of aspect 894, wherein said monocytes or monocytic progenitors are cultured with said stem cells at a ratio of 1 monocyte or monocytic progenitor cell to 1 stem cells.

[0902] 896. The method of aspect 851, wherein said monocytes or monocytic progenitors are cultured with said stem cells for a time period of 1 hour to 7 days.

[0903] 897. The method of aspect 896, wherein said monocytes or monocytic progenitors are cultured with said stem cells for a time period of 12 hours to 5 days.

[0904] 898. The method of aspect 897, wherein said monocytes or monocytic progenitors are cultured with said stem cells for a time period of 1 to 3 days.

[0905] 899. The method of aspect 851, wherein said monocytes or monocytic progenitors are cultured with said fibroblasts in a media selected from a group comprising of: Roswell Park Memorial Institute (RPMI-1640), Dublecco's Modified Essential Media (DMEM), Dublecco's Modified Essential Media - Low Glucose (DMEM- LG), Eagle's Modified Essential Media (EMEM), Optimem, Iscove's Media, or combinations thereof.

[0906] 900. The method of aspect 899, wherein said media is supplemented with growth factors selected from a group comprising of: human platelet rich plasma, platelet lysate, umbilical cord blood serum, autologous serum, human serum, serum replacement, or combinations thereof.

[0907] 901. The method of aspect 851, wherein in said therapeutic macrophages are capable of stimulating growth of new blood vessels.

[0908] 902. The method of aspect 851, wherein in said therapeutic macrophages areM2 macrophage.

[0909] 903. The method of aspect 851, wherein said stem cells are derived from the perivascular areas of Wharton's Jelly.

[0910] 904. The method of aspect 851, wherein said stem cells are exposed to hyperthermia to enhance therapeutic activity.

[0911] 905. The method of aspect 851, wherein said therapeutic macrophages are neuroregenerative.

[0912] 906. The method of aspect 905, wherein said neuroregenerative macrophages are capable of protecting neurons from apoptosis.

[0913] 907. The method of aspect 905, wherein said neuroregenerative macrophages are capable of protecting neurons from excitotoxicity.

[0914] 908. The method of aspect 905, wherein said neuroregenerative macrophages are capable of maintaining neuronal connections.

[0915] 909. The method of aspect 905, wherein said neuroregenerative macrophages are capable of suppressing microglial activation.

[0916] 910. The method of aspect 905, wherein said neuroregenerative macrophages are capable of stimulating proliferation of endogenous neural progenitor cells.

[0917] 911. The method of aspect 851, wherein said therapeutic macrophages are hepatoprotective.

[0918] 912. The method of aspect 911, wherein said hepatoprotective macrophages induce liver regeneration.

[0919] 913. The method of aspect 911, wherein said hepatoprotective macrophages suppress liver fibrosis.

[0920] 914. The method of aspect 911, wherein said hepatoprotective macrophages inhibit production of TGF-beta.

[0921] 915. The method of aspect 911, wherein said hepatoprotective macrophages inhibit activation of the SMAD pathway.

[0922] 916. The method of aspect 911, wherein said hepatoprotective macrophages suppress oxidative stress.

[0923] 917. The method of aspect 911, wherein said hepatoprotective macrophages stimulate hepatic angiogenesis.

[0924] 918. The method of aspect 851, wherein said therapeutic macrophages are renoprotective.

[0925] 919. The method of aspect 851, wherein said therapeutic macrophages are cardioprotective.

[0926] 920. The method of aspect 919, wherein said cardioprotective macrophages preserve viability of cardiomyocytes after an infarct.

[0927] 921. The method of aspect 919, wherein said cardioprotective macrophages preserve viability of cardiac progenitor cells.

[0928] 922. The method of aspect 919, wherein said cardioprotective macrophages preserve viability of cardiac endothelial cells after an infarct.

[0929] 923. The method of aspect 919, wherein said cardioprotective macrophages reduce cardiac fibrosis.

[0930] 924. The method of aspect 919, wherein said cardioprotective macrophages inhibit pathological cardiac remodeling.

[0931] 925. The method of aspect 919, wherein said cardioprotective macrophages suppress oxidative stress in cardiac tissue.

[0932] 926. The method of aspect 919, wherein said cardioprotective macrophages recruit regenerative cells into the cardiac tissue.

[0933] 927. The method of aspect 926, wherein said regenerative cells are endogenous cardiac progenitor cells.

[0934] 928. The method of aspect 927, wherein said endogenous cardiac progenitor cells express c-kit.

[0935] 929. The method of aspect 927, wherein said endogenous cardiac progenitor cells express CD133.

[0936] 930. The method of aspect 927, wherein said endogenous cardiac progenitor cells possess ability to efflux rhodamine 231.

[0937] 931. The method of aspect 926, wherein said regenerative cells are bone marrow regenerative cells.

[0938] 932. The method of aspect 931, wherein said bone marrow regenerative cells express the marker CD34.

[0939] 933. The method of aspect 931, wherein said bone marrow regenerative cells are endothelial progenitor cells.

[0940] 934. The method of aspect 933, wherein said endothelial progenitor cells are capable of healing damaged blood vessels.

[0941] 935. The method of aspect 934, wherein said damaged blood vessel is an atherosclerotic blood vessel.

[0942] 936. The method of aspect 934, wherein said damaged blood vessel is infarct associated blood vessel.

[0943] 937. The method of aspect 934, wherein said damaged blood vessel is a blood vessel that has been exposed to disseminated intravascular coagulation.

[0944] 938. The method of aspect 933, wherein said endothelial progenitor cells are capable of forming endothelial colonies when plated on fibronectin.

[0945] 939. The method of aspect 933, wherein said endothelial progenitor cells possess CXCR4.

[0946] 940. The method of aspect 933, wherein said endothelial progenitor cells possess tie-2.

[0947] 941. The method of aspect 933, wherein said endothelial progenitor cells possess VE Cadherin.

[0948] 942. The method of aspect 933, wherein said endothelial progenitor cells possess CD31.

[0949] 943. The method of aspect 933, wherein said endothelial progenitor cells possess CD34.

[0950] 944. The method of aspect 933, wherein said endothelial progenitor cells possess CD133.

[0951] 945. The method of aspect 933, wherein said endothelial progenitor cells possess CD117.

[0952] 946. The method of aspect 933, wherein said endothelial progenitor cells possess CD105.

[0953] 947. The method of aspect 933, wherein said endothelial progenitor cells possess ETV2.

[0954] 948. The method of aspect 933, wherein said endothelial progenitor cells possess CD146.

[0955] 949. The method of aspect 933, wherein said endothelial progenitor cells possess flk-1.

[0956] 950. The method of aspect 933, wherein said endothelial progenitor cells possess flt-4.

[0957] 951. The method of aspect 933, wherein said bone marrow regenerative cell is a hematopoietic stem cell.

[0958] 952. The method of aspect 951, wherein said hematopoietic stem cell possessesIL-3 receptor.

[0959] 953. The method of aspect 951, wherein said hematopoietic stem cell possessesIL-6 receptor.

[0960] 954. The method of aspect 951, wherein said hematopoietic stem cell possesses thrombopoietin receptor.

[0961] 955. The method of aspect 951, wherein said hematopoietic stem cell expressesCD34.

[0962] 956. The method of aspect 951, wherein said hematopoietic stem cell expressesCD133.

[0963] 957. The method of aspect 951, wherein said hematopoietic stem cell does not express CD38.

[0964] 958. The method of aspect 851, wherein said umbilical cord blood stem cell expresses CD56.

[0965] 959. The method of aspect 851, wherein said umbilical cord blood stem cell expresses CD56.

[0966] 960. The method of aspect 851, wherein said umbilical cord blood stem cell expresses dopamine D3 receptor.

[0967] 961. The method of aspect 851, wherein said umbilical cord blood stem cell expresses dopamine D3 receptor.

[0968] 962. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Notch.

[0969] 963. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Notch 1.

[0970] 964. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Jagged-1.

[0971] 965. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Delta.

[0972] 966. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Delta-1.

[0973] 967. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Delta-4.

[0974] 968. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Oct-3 / 4.

[0975] 969. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Rex-1.

[0976] 970. The method of aspect 851, wherein said umbilical cord blood stem cell expresses Nanog.

[0977] 971. The method of aspect 851, wherein said umbilical cord blood stem cell expresses LIF-STAT2.

[0978] 972. The method of aspect 851, wherein said umbilical cord blood stem cell expresses STAT5.

[0979] 973. The method of aspect 851, wherein said umbilical cord blood stem cell expresses STAT5A.

[0980] 974. The method of aspect 851, wherein said umbilical cord blood stem cell expresses sonic hedgehog.

[0981] 975. The method of aspect 851, wherein said umbilical cord blood stem cell expresses BMP2.

[0982] 976. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of one or more selected from a group comprising of: Wntl, Wnt2, Wnt2b / 13, Wnt3, Wnt3a, Wnt4, Wnt5a, Wnt5b, Wnt6, Wnt7a, Wnt7b, Wnt7c, Wnt8, Wnt8a, Wnt8b, Wnt8c, WntlOa, WntlOb, Wntll, Wntl4, Wntl5, or Wntl6.

[0983] 977. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Notch.

[0984] 978. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Delta.

[0985] 979. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Serrate.

[0986] 980. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Jagged.

[0987] 981. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Mastermind.

[0988] 982. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Enhancer of Split.

[0989] 983. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Hesl.

[0990] 984. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Hairless.

[0991] 985. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Suppressor Hairless.

[0992] 986. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of RBP-Jk.

[0993] 986. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Desert hedgehog.

[0994] 987. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Sonic hedgehog.

[0995] 988. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Indian hedgehog.

[0996] 989. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Gli.

[0997] 990. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Gli-1.

[0998] 991. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Gli-3.

[0999] 992. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Patched.

[1000] 993. The method of aspect 851, wherein said umbilical cord blood stem cells are treated with an agonist of Patched-1.

[1001] 994. A method of generating mesenchymal stem cells by treatment of pluripotent stem cells with one or more inhibitors of TGF-beta together with an inhibitor of histone deacetylase.

[1002] 995. The method of aspect 994, wherein said mesenchymal stem cell expresses c-kit.

[1003] 996. The method of aspect 994, wherein said inhibitor of histone deacetylase is valproic acid.

[1004] 997. The method of aspect 994, wherein said inhibitor of histone deacetylase is phenylbutyrate.

[1005] 998. The method of aspect 994, wherein said inhibitor of histone deacetylase is trichostatin A.

[1006] 999. The method of aspect 994, wherein said inhibitor of histone deacetylase is sulforaphane.

[1007] 1000. The method of aspect 994, wherein said TGF-beta inhibitor is an inhibitor of SMAD2.

[1008] 1001. The method of aspect 994, wherein said TGF-beta inhibitor is an inhibitor of SMAD4BRIEF DESCRI PTION OF DRAWINGS

[1009] Figure 1 is a bar graph showing the results of CD015 expression in generated mesenchymal stem cells in relation to the amount of bone matrix used.DETAILED DESCRIPTION OF TH E I NVENTION

[1010] The current invention teaches subject matter relates to therapeutic cells in the family of mesenchymal stem cells, or MSCs, including human mesenchymal stem cells. More particularly, this invention provides methods of generating mesenchymal stem cells from pluripotent stem cell such as induced pluripotent stem cells (iPSCs), wherein the iPSCs are cultured under conditions to produce mesenchymal stem cells. Prior to being cultured under conditions to produce mesenchymal stem cells, the iPSCs may be genetically engineered with at least one polynucleotide encoding at least one biologically active protein or polypeptide or biologically active fragment, derivative, or analogue thereof, thus enabling one to produce genetically engineered mesenchymal stem cells from the genetically engineered iPSCs that express sustained levels of the at least one biologically active protein or polypeptide, or biologically active fragment, derivative, or analogue thereof. Said molecules may be useful in endowing saidproduced mesenchymal stem cells with therapeutic properties such as: a) enhanced angiogenesis; b) enhanced neurogenesis; c) enhanced ability to stimulate endogenous progenitor cells and d) enhanced viability.

[1011] Various terms are used to describe cells in culture. Cell culture refers generally to cells taken from a living organism and grown under controlled condition ("in culture" or "cultured"). A primary cell culture is a culture of cells, tissues, or organs taken directly from an organism(s) before the first subculture. Cells are expanded in culture when they are placed in a growth medium under conditions that facilitate cell growth and / or division, resulting in a larger population of the cells. When cells are expanded in culture, the rate of cell proliferation is sometimes measured by the amount of time needed for the cells to double in number. This is referred to as doubling time.

[1012] A cell line is a population of cells formed by one or more subcultivations of a primary cell culture. Each round of subculturing is referred to as a passage. When cells are subcultured, they are referred to as having been passaged. A specific population of cells, or a cell line, is sometimes referred to or characterized by the number of times it has been passaged. For example, a cultured cell population that has been passaged ten times may be referred to as a PIO culture. The primary culture, i.e., the first culture following the isolation of cells from tissue, is designated P0. Following the first subculture, the cells are described as a secondary culture (Pl or passage 1). After the second subculture, the cells become a tertiary culture (P2 or passage 2), and so on. It will be understood by those of skill in the art that there may be many population doublings during the period of passaging; therefore the number of population doublings of a culture is greater than the passage number. The expansion of cells (i.e., the number of population doublings) during the period between passaging depends on many factors, including but not limited to the seeding density, substrate, medium, growth conditions, and time between passaging.

[1013] A conditioned medium is a medium in which a specific cell or population of cells has been cultured, and then removed. When cells are cultured in a medium, they may secrete cellular factors that can provide trophic support to other cells. Such trophic factors include, but are not limited to hormones, cytokines, extracellular matrix (ECM), proteins, vesicles, antibodies, and granules. The medium containing the cellular factors is the conditioned medium. In some embodiments the invention teaches the use of conditioned media, or concentrated conditioned media, or exosomes isolated from conditioned media of EPC or MSC to promote tolerogenesis.

[1014] As used herein, the term Growth Medium generally refers to a medium sufficient for the culturing of umbilicus-derived cells. In particular, one presently preferred medium for the culturing of the cells of the invention herein comprises Dulbecco's Modified Essential Media (also abbreviated DMEM herein). Particularly preferred is DMEM-low glucose (also DMEM-LG herein) (Invitrogen, Carlsbad, Calif.). The DMEM-low glucose is preferably supplemented with 15% (v / v) fetal bovine serum (e.g. defined fetal bovine serum, Hyclone, Logan Utah), antibiotics / antimycotics (preferably penicillin (100 Units / milliliter), streptomycin (100 milligrams / m illiliter), and amphotericin B (0.25 micrograms / milliliter), (Invitrogen, Carlsbad, Calif.)), and 0.001% (v / v) 2-mercaptoethanol (Sigma, St. Louis Mo.). In some cases different growth media are used, or different supplementations are provided, and these are normally indicated in the text as supplementations to Growth Medium.

[1015] Also relating to the present invention, the term standard growth conditions, as used herein refers to culturing of cells at 37. degree. C., in a standard atmosphere comprising 5% CO. sub.2. Relative humidity is maintained at about 100%. While foregoing the conditions are useful for culturing, it is to be understood that such conditions are capable of being varied by the skilled artisan who will appreciate the options available in the art for culturing cells, for example, varying the temperature, CO. sub.2, relative humidity, oxygen, growth medium, and the like.

[1016] "Mesenchymal stem cell" or "MSC" in some embodiments refers to cells that are (1) adherent to plastic, (2) express CD73, CD90, and CD105 antigens, while being CD14, CD34, CD45, and HLA-DR negative, and (3) possess ability to differentiate to osteogenic, chondrogenic and adipogenic lineage. Other cells possessing mesenchymal-like properties are included within the definition of "mesenchymal stem cell", with the condition that said cells possess at least one of the following: a) regenerative activity; b) production of growth factors; c) ability to induce a healing response, either directly, or through elicitation of endogenous host repair mechanisms. As used herein, "mesenchymal stromal cell" or ore mesenchymal stem cell can be used interchangeably. Said MSCcan be derived from any tissue including, but not limited to, bone marrow, adipose tissue, amniotic fluid, endometrium, trophoblast-derived tissues, cord blood, Wharton jelly, placenta, amniotic tissue, derived from pluripotent stem cells, and tooth. In some definitions of "MSC", said cells include cells that are CD34 positive upon initial isolation from tissue but are similar to cells described about phenotypically and functionally. As used herein, "MSC" may includes cells that are isolated from tissuesusing cell surface markers selected from the list comprised of NGF-R, PDGF-R, EGF-R, IGF-R, CD29, CD49a, CD56, CD63, CD73, CD105, CD106, CD140b, CD146, CD271, MSCA-1, SSEA4, STRO-1 and STRO-3 or any combination thereof, and satisfy the ISCT criteria either before or after expansion. Furthermore, as used herein, in some contexts, "MSC" includes cells described in the literature as bone marrow stromal stem cells (BMSSC), marrow-isolated adult multipotent inducible cells (MIAMI) cells, multipotent adult progenitor cells (MAPC), mesenchymal adult stem cells (MASCS), MultiStem®, Prochymal®, remestemcel-L, Mesenchymal Precursor Cells (MPCs), Dental Pulp Stem Cells (DPSCs), PLX cells, PLX-PAD, AlloStem®, Astrostem®, Ixmyelocel-T, MSC-NTF, NurOwn™, Stemedyne™-MSC, Stempeucel®, StempeucelCLI, StempeucelOA, HiQCell, Hearticellgram-AMI, Revascor®, Cardiorel®, Cartistem®, Pneumostem®, Promostem®, Homeo-GH, AC607, PDA001, SB623, CX601, AC607, Endometrial Regenerative Cells (ERC), adipose-derived stem and regenerative cells (ADRCs).

[1017] Oct -4 (oct-3 in humans) is a transcription factor expressed in the pregastrulation embryo, early cleavage stage embryo, cells of the inner cell mass of the blastocyst, and embryonic carcinoma ("EC") cells (Nichols, J. et al. (1998) Cell 95: 379-91), and is down- regulated when cells are induced to differentiate. The oct -4 gene (oct-3 in humans) is transcribed into at least two splice variants in humans, oct-3A and oct-3B. The oct-3B splice variant is found in many differentiated cells whereas the oct-3A splice variant (also previously designated oct-3 / 4) is reported to be specific for the undifferentiated embryonic stem cell. See Shimozaki et al. (2003) Development 130: 2505-12. Expression of oct-3 / 4 plays an important role in determining early steps in embryogenesis and differentiation. Oct-3 / 4, in combination with rox-1, causes transcriptional activation of the Zn-finger protein rex-1, which is also required for maintaining ES cells in an undifferentiated state (Rosfjord, E. and Rizzino, A. (1997) Biochem Biophys Res Commun 203: 1795-802; Ben-Shushan, E. et al. (1998) Mol Cell Biol 18: 1866-78). In some embodiments of the invention mesenchymal stem cells are selected for pluripotent derived

[1018] expression of OCT-4. In other embodiments, OCT-4 expression is used as a means of identifying cells for culture and expansion subsequent to exposure to various culture conditions.

[1019] Presently preferred are methods which provide cells which require no exogenous growth factors, except as are available in the supplemental serum provided with the Growth Medium. Also provided herein are methods of deriving umbilical cellscapable of expansion in the absence of particular growth factors. The methods are similar to the method above, however they require that the particular growth factors (for which the cells have no requirement) be absent in the culture medium in which the cells are ultimately resuspended and grown in. In this sense, the method is selective for those cells capable of division in the absence of the particular growth factors. Preferred cells in some embodiments are capable of growth and expansion in chemically-defined growth media with no serum added. In such cases, the cells may require certain growth factors, which can be added to the medium to support and sustain the cells. Presently preferred factors to be added for growth on serum-free media include one or more of FGF, EGF, IGF, and PDGF. In more preferred embodiments, two, three or all four of the factors are add to serum free or chemically defined media. In other embodiments, LIF is added to serum-free medium to support or improve growth of the cells.

[1020] Also provided are methods wherein the cells can expand in the presence of from about 5% to about 20% oxygen in their atmosphere. Methods to obtain cells that require L-valine require that cells be cultured in the presence of L-valine. After a cell is obtained, its need for L-valine can be tested and confirmed by growing on D-valine containing medium that lacks the L-isomer. Methods are provided wherein the cells can undergo at least 25, 30, 35, or 40 doublings prior to reaching a senescent state. Methods for deriving cells capable of doubling to reach 10.sup.14 cells or more are provided.Preferred are those methods which derive cells that can double sufficiently to produce at least about 10. sup.14, 10.sup.15, 10.sup.16, or 10.sup.17 or more cells when seeded at from about 10.sup.3 to about 10.sup.6 cells / cm.sup.2 in culture. Preferably these cell numbers are produced within 80, 70, or 60 days or less. In one embodiment, pluripotent derived mesenchymal stem cells are isolated and expanded, and possess one or more markers selected from a group comprising of CD10, CD13, CD44, CD73, CD90, CD141, PDGFr-alpha, or HLA-A,B,C. In addition, the cells do not produce one or more of CD31, CD45, CD117, CD141, or HLA-DR,DP, DQ.

[1021] In one embodiment generation of mesenchymal stem cells from pluripotent stem cells is accomplished by administration of a TGF-beta inhibitor together with hepatocyte growth factor. One of skill in the art may utilize various inhibitors of the cytokine itself or of signaling pathways associated with said TGF-beta. For example, in one embodiment of the invention TGF-beta inhibition is accomplished by administration of a smad inhibitor.

[1022] In a non-limiting embodiment, induced pluripotent stem cells are cultured in a medium, such as the feeder-free medium mTeSRl (STEMCELL Technologies) that has been supplemented with a TGF-P inhibitor such as SB431542 in an atmosphere containing 7.5 vol. % CO2 for 25 days. The cells then are transfered to a tissue culture plastic dish having a hydrophilic surface, and which contains a medium, such as a modified human ES-MSC medium containing knockout serum replacement, nonessential amino acids, antibiotic such as penicillin and streptomycin, glutamine, - mercaptoethanol, and bFGF, which has been supplemented with a TGF-P inhibitor such as SB-431542. The medium is changed daily, and the cells are passaged at 80%-90% confluence about every 3 days. The cells are cultured for a total of about 21 days to provide a majority of cells that are positive for MSC surface markers. Such mesenchymal stem cells also are known as iPSC-MSCs. The iPSC-MSCs then can be cultured in the presence of a standard medium, such as 20% fetal bovine serum (FBS) a-MEM medium, and then harvested for further experiments or for use in treating diseases or disorders, or for regenerating cells, tissues, or organs.

[1023] The mesenchymal stem cells formed from the induced pluripotent stem cells in accordance with the present invention thus have several desirable properties and characteristics that make the mesenchymal stem cells more stable, and whereby such mesenchymal stem cells are less likely to form or cause tumors, cancers, or teratomas, and thus are more desirable for use in therapy than other mesenchymal stem cells. Thus, in accordance with another aspect of the present invention, there are provided isolated human mesenchymal stem cells derived from human induced pluripotent stem cells that express no more than 1% of the levels of the Nanog, Oct. 4, Ecad, and Foxa2 genes than the induced pluripotent stem cells from which the mesenchymal stem cells were derived. In a non-limiting embodiment, the isolated human mesenchymal stem cells are at least 95% positive for the epitopes CD73, CD105, and CD166. In another nonlimiting embodiment, the isolated human mesenchymal stem cells are at least 85% positive for the epitopes CD44 and CD90. In yet another non-limiting embodiment, the isolated human mesenchymal stem cells are no more than 5% positive for the epitopes HLA-DR, CDllb, CD24, CD34, and CD45. Furthermore, the isolated human mesenchymal stem cells of the present invention, in a non-limiting embodiment, contain the following levels of messenger RNAs (mRNAs) relative to a standardized preparation of MSC obtained from bone marrow (Sample No. 7075, available from the Institute for Regenerative Medicine, Texas A&M College of Medicine, Temple, Texas 76502): about80% to about 120% of the mesodermal marker CD140A, about 550% to about 650% of the angiogenic gene VEGF, and less than 20% ±5% of the following genes known to promote the growth and metastasis of cancer cells: I LR1, mPGESl, IL-6, TGF-13R2, I D3, SDF1, HAS1, and HAS2. The isolated human mesenchymal stem cells of the present invention also stop dividing in culture after 70 to 100 population doublings under conditions in which MSCs obtained from bone marrow also stop dividing and therefore are less likely than immortal cells to produce tumors or cancers in patients, and are less likely to form teratomas in culture or tumors after injection into immunodeficient animals, such as immunodeficient mice. The isolated human mesenchymal stem cells of the present invention may be administered in an amount effective to treat a variety of diseases and disorders, and to regenerate a variety of cells, tissues, and organs. The isolated human mesenchymal stem cells may be administered systematically such as by intramuscular, intravenous, intraperitoneal or intra-arterial administration or may be administered directly to an affected cell, tissue, or organ. The isolated human mesenchymal stem cells may be administered in conjunction with an acceptable pharmaceutical carrier adjuvant or excipient known to those skilled in the art. Such diseases and disorders include, but are not limited to, inflammatory diseases, disorders, and conditions, eye diseases and disorders, such as macular degeneration, diseases of the cornea, eye injuries, including corneal injuries, cardiac disease, including myocardial infarction, brain injury, brain trauma, brain diseases and disorders, including stroke and Alzheimer's disease, neuro motor diseases such as Parkinson's Disease, autoimmune diseases, including diabetes, obesity, and tumors, including malignant and non- malignant tumors. Cells, tissues, or organs which may be regenerated in accordance with the isolated human mesenchymal stem cells of the present invention include, but are not limited to, bone tissue, eye tissue, including corneal tissue, cardiac tissue, including cardiac muscle and the coronary arteries, as well as any other cell, tissue or organ known to be regenerated by mesenchymal stem cells. The exact dosage of mesenchymal stem cells to be administered is dependent upon a variety of factors, including but not limited to the age, weight, height, and sex of the patient, the disease or disorder being treated, and the extent and severity thereof, or the cells, tissue, or organ to be regenerated. It is to be understood, however, that the scope of the present invention is not intended to be limited to the treatment of any particular disease, condition, or disorder, or to the regeneration of any particular cell, tissue, or organ. The isolated human mesenchymal stem cells of the present invention, prepared ashereinabove described, and having the properties hereinabove described, may be genetically engineered with at least one polynucleotide encoding at least one biologically active protein or polypeptide or biologically active fragment, derivative, or analogue thereof. Thus, in accordance with an aspect of the present invention, there is provided a method of producing genetically engineered mesenchymal stem cells from induced pluripotent stem cells. The method comprises introducing into the induced pluripotent stem cells at least one polynucleotide encoding at least one biologically active protein or polypeptide, or biologically active fragment, analogue, or derivative thereof to provide genetically engineered induced pluripotent stem cells. The genetically engineered pluripotent stem cells then are cultured as hereinabove described to produce genetically engineered mesenchymal stem cells, such as mammalian mesenchymal stem cells. In a non-limiting embodiment, the genetically engineered mammalian mesenchymal stem cells are primate mesenchymal stem cells, including human mesenchymal stem cells. Thus, the genetically engineered induced pluripotent stem cells are cultured in a medium containing a TGF-P inhibitor and in an atmosphere containing from about 7 vol. % to about 8 vol. % CO2 (about 7.5 vol. % CO2 in another non-limiting embodiment) for a period of time of from about 20 days to about 35 days (about 25 days in another non-limiting embodiment). The genetically engineered cells then are transferred to a culture dish having a hydrophilic surface, such as those hereinabove described, and cultured in a medium containing a TGF-P inhibitor for a period of time (in a non-limiting embodiment, 21 days) sufficient to produce genetically engineered mesenchymal stem cells. The at least one polynucleotide including at least one biologically active protein or polypeptide or biologically active fragment or derivative may be in the form of DNA (including but not limited to genomic DNA (gDNA) or cDNA, or RNA. The at least one polynucleotide encoding at least one biologically active protein or polypeptide or biologically active fragment, derivative, or analogue thereof may be contained in an appropriate expression vector, such as an adenoviral vector, adeno-associated virus vector, retroviral vector, or lentiviral vector that is introduced into the induced pluripotent stem cells, or may be contained in a transposon that is introduced into the cell, or the at least one polynucleotide may be introduced into the cell as naked DNA or RNA. Such introduction of the at least one polynucleotide may be introduced into the cell by any of a variety of means known to those skilled in the art, such as calcium phosphate precipitation, liposomes, gene guns, or by clustered regularly interspersed short palindromic repeats, or CRISPR, technology.

[1024] Biologically active proteins or polypeptides, or biologically active fragments, derivatives, or analogues thereof that may be introduced into the induced pluripotent stem cells, prior to the production of mesenchymal stem cells therefrom, include polynucleotides encoding various therapeutic agents including, but not limited to, antiinflammatory or inflammation modulatory agents, such as TSG-6, anti-angiogenic agents, tumor necrosis factors, interleukins, growth factors, anti-clotting agents, bone morphogenic proteins (BMPs), such as BMP-2, hormones, such as insulin, anti-tumor agents, and negative selective markers. It is to be understood, however, that the scope of the present invention is not intended to be limited to any particular biologically active protein or polypeptide, or biologically active fragment, derivative, or analogue thereof. In a non-limiting embodiment the at least one biologically active protein or polypeptide or biologically active fragment, derivative, or analogue is tumor necrosis factor alpha stimulating gene 6 (TSG-6) protein or a biologically active fragment, derivative, or analogue thereof.

[1025] The optimal dose of pluripotent derived MSC for some embodiments will be in the range of doses used for autologous, mononuclear bone marrow transplantation. For fairly pure preparations of pluripotent derived MSC, optimal doses in various embodiments will range from 10.sup.4 to 10.sup.8 pluripotent derived MSC cells / kg of recipient mass per administration. In some embodiments the optimal dose per administration will be between 10.sup.5 to 10. sup.7 pluripotent derived MSC cells / kg. In many embodiments the optimal dose per administration will be 5.times.l0.sup.5 to 5. times.10. sup.6 pluripotent derived MSC cells / kg. By way of reference, higher doses in the foregoing are analogous to the doses of nucleated cells used in autologous mononuclear bone marrow transplantation. Some of the lower doses are analogous to the number of CD34.sup.+ cells / kg used in autologous mononuclear bone marrow transplantation.

[1026] It is to be appreciated that a single dose may be delivered all at once, fractionally, or continuously over a period of time. The entire dose also may be delivered to a single location or spread fractionally over several locations. In various embodiments, pluripotent derived MSC may be administered in an initial dose, and thereafter maintained by further administration of pluripotent derived MSC. pluripotent derived MSC may be administered by one method initially, and thereafter administered by the same method or one or more different methods. The subject's MSC levels can be maintained by the ongoing administration of the cells. Various embodiments administerthe pluripotent derived MSC either initially or to maintain their level in the subject or both by intravenous injection. In a variety of embodiments, other forms of administration, are used, dependent upon the patient's condition and other factors, discussed elsewhere herein. It is noted that human subjects are treated generally longer than experimental animals; but, treatment generally has a length proportional to the length of the disease process and the effectiveness of the treatment. Those skilled in the art will take this into account in using the results of other procedures carried out in humans and / or in animals, such as rats, mice, non-human primates, and the like, to determine appropriate doses for humans. Such determinations, based on these considerations and taking into account guidance provided by the present disclosure and the prior art will enable the skilled artisan to do so without undue experimentation. Suitable regimens for initial administration and further doses or for sequential administrations may all be the same or may be variable. Appropriate regiments can be ascertained by the skilled artisan, from this disclosure, the documents cited herein, and the knowledge in the art.

[1027] The dose, frequency, and duration of treatment will depend on many factors, including the nature of the disease, the subject, and other therapies that may be administered. Accordingly, a wide variety of regimens may be used to administer MSC. In some embodiments MSC are administered to a subject in one dose. In others MSC are administered to a subject in a series of two or more doses in succession. In some other embodiments wherein MSC are administered in a single dose, in two doses, and / or more than two doses, the doses may be the same or different, and they are administered with equal or with unequal intervals between them. MSC may be administered in many frequencies over a wide range of times. In some embodiments, pluripotent derived MSC are administered over a period of less than one day. In other embodiment they are administered over two, three, four, five, or six days. In some embodiments pluripotent derived MSC are administered one or more times per week, over a period of weeks. In other embodiments they are administered over a period of weeks for one to several months. In various embodiments they may be administered over a period of months. In others they may be administered over a period of one or more years. Generally lengths of treatment will be proportional to the length of the disease process, the effectiveness of the therapies being applied, and the condition and response of the subject being treated.

[1028] The induced pluripotent stem cell may be produced by expressing or inducing the expression of one or more reprogramming factors in a somatic cell. The somatic cell is a fibroblast, such as a dermal fibroblast, synovial fibroblast, or lung fibroblast, or a non-fibroblastic somatic cell. The somatic cell is reprogrammed by expressing at least 1, 2, 3, 4, 5 reprogramming factors. The reprogramming factors may be selected from Oct 3 / 4, Sox2, NANOG, Lin28, c Myc, and Klf4. Expression of the reprogramming factors may be induced by contacting the somatic cells with at least one agent, such as a small organic molecule agents, that induce expression of reprogramming factors.

[1029] The somatic cell may also be reprogrammed using a combinatorial approach wherein the reprogramming factor is expressed (e.g., using a viral vector, plasmid, and the like) and the expression of the reprogramming factor is induced (e.g., using a small organic molecule.) For example, reprogramming factors may be expressed in the somatic cell by infection using a viral vector, such as a retroviral vector or a lentiviral vector. Also, reprogramming factors may be expressed in the somatic cell using a non- integrative vector, such as an episomal plasmid. See, e.g., Yu et al., Science. 2009 May 8; 324(5928):797-801, which is hereby incorporated by reference in its entirety. When reprogramming factors are expressed using non-integrative vectors, the factors may be expressed in the cells using electroporation, transfection, or transformation of the somatic cells with the vectors. For example, in mouse cells, expression of four factors (Oct3 / 4, Sox2, c myc, and Klf4) using integrative viral vectors can be used to reprogram a somatic cell. In human cells, expression of four factors (Oct34, Sox2, NANOG, and Lin28) using integrative viral vectors can be used to reprogram a somatic cell.

[1030] Once the reprogramming factors are expressed in the cells, the cells may be cultured. Over time, cells with ES characteristics appear in the culture dish. The cells may be chosen and subcultured based on, for example, ES morphology, or based on expression of a selectable or detectable marker. The cells may be cultured to produce a culture of cells that resemble ES cells-these are putative iPS cells. iPS cells typically can be identified by expression of the same markers as other embryonic stem cells, though a particular iPS cell line may vary in its expression profile. Exemplary iPS cells may express Oct -4, alkaline phosphatase, SSEA 3 surface antigen, SSEA 4 surface antigen, TRA 1 60, anchor TRA 1 81.

[1031] The invention provides for generation of regenerative T cells through differentiation of pluripotent stem cells in the presence of mesenchymal stem cells orproducts derived thereof. Said products include exosomes, microvesicles and apoptotic bodies.

[1032] According to a preferred embodiment, the pluripotent stem cells are cultured under conditions enabling to induce the formation of embryoid bodies. For this purpose, cell culture can be performed, for illustrative purposes, in a low-adhesion plate (Sigma- Aldrich, Fisher), which favors the appearance of cell aggregates in three dimensions (embryoid bodies) and reproduces in a more efficient way the intercellular interactions existing during the development of the embryo in the body of the animal.

[1033] In an embodiment, the culture of pluripotent stem cells is performed in a culture medium suitable to induce the formation of embryoid bodies, for at least 9 days, under conditions allowing to obtain embryoid bodies comprising at least 5% of CD34+CD43+ cells. The invention teaches that this step may be performed in the presence of interferon gamma activated mesenchymal stem cells, preferably at a ratio of one to one. Culture media suitable for the growth of mesenchymal stem cells are known from the state of the art. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises a serum-free culture medium suitable for the growth of mesenchymal stem cells, for example the StemPro-34 SFM medium (ThermoFisher). According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 0.1 to about 5% of L-glutamine, preferably about 1% of L-glutamine. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 0.1 to about 5% of non-essential amino acids, preferably about 1% of non-essential amino acids. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 0.01 to about 0.5% of 2-mercaptoethanol, preferably about 0.1% of 2-mercaptoethanol. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 10 to about 1000 U / mL of penicillin, preferably about 100 U / mL of penicillin. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 10 to about 1000 ng / mL of streptomycin, preferably about 100 ng / mL of streptomycin. According to an embodiment, the culture medium to induce the formation of embryoid bodies comprises from about 5 to about 1000 .mu.g / mL, preferably about 50 .mu.g / mL of ascorbic acid. According to an embodiment, a serum-free culture medium suitable for the growth of mesenchymal stem cells, for example StemPro-34 SFM medium, comprises L-glutamine, non-essential amino acids, 2-mercaptoethanol, penicillin,streptomycin and / or ascorbic acid as described above. In an embodiment, the culture of the pluripotent stem cells in step a) is carried out in a serum-free culture medium suitable for the growth of mesenchymal stem cells, for example StemPro-34 SFM medium (ThermoFisher), comprising BMP, FGF2, VEGF, SCF, Flt3-L and / or IL-3, for at least 9 days, under conditions allowing to obtain embryoid bodies comprising at least 5% of CD34+CD43+ cells. In an embodiment, the culture of the pluripotent stem cells in step a) is carried out in a serum-free culture medium suitable for the growth of mesenchymal stem cells, for example StemPro-34 SFM medium (ThermoFisher), comprising BMP, FGF2, VEGF, SCF, Flt3-L and IL-3, for at least 9 days, under conditions allowing to obtain embryoid bodies comprising at least 5% of CD34+CD43+ cells. According to a preferred embodiment, the pluripotent stem cells are first incubated in the presence of BMP (bone morphogenetic protein) to facilitate the induction of the formation of embryoid bodies. According to a preferred embodiment, the pluripotent stem cells are incubated for 10 to 48 hours, preferably for one day, in the presence of BMP. According to another embodiment, the cells are incubated in the presence of 3 to 300 ng / mL of BMP, preferably from 10 to 100 ng / mL of BMP, more preferably from 20 to 50 ng / mL of BMP, and particularly about 30 ng / mL of BMP. Preferably, the BMP is BMP-4, more preferably human BMP-4 (hBMP-4). More preferably, the cells are incubated for one day in the presence of about 30 ng / mL of hBMP-4a, preferably on DO (start of the second phase). In an embodiment, after the incubation in the presence of BMP, preferably BMP-4, a mixture comprising BMP (preferably BMP-4) and FGF-2 is added to the medium to allow induction of mesoderm. Preferably between about 3 ng / mL and about 300 ng / mL of BMP, preferably BMP-4, and between about 0.5 ng / mL and about 50 ng / mL of FGF2 is added to the medium, preferably about 30 ng / mL of BMP, preferably BMP-4, and about 5 ng / mL of FGF2 are added to the medium. According to an embodiment, this addition is carried out all at once, preferably on day 1 of the phase of induction of the formation of embryoid bodies (day DI). In an embodiment, a solution comprising growth factors and / or cytokines is added every two days, preferably from day 3 of the phase of induction of the formation of embryoid bodies (day D3), until the end of the induction phase of the formation of embryoid bodies. The end of the induction phase of the formation of embryoid bodies corresponds to the moment when the embryoid bodies are dissociated. This can for example occur on day 5, day 6, day 7, day 8, day 9, day 10, day 11, day 12 of the induction phase of the formation of the embryoid bodies, or later (D5, D6, D7, D8, D9, D10, Dll, D12 or later). In a particular embodiment, step a) ofculture of the pluripotent stem cells is carried out for at least 9 days. In an embodiment, step a) is carried out for 9 to 17 days, preferably for 9 to 15 days, preferably for 9 to 14 days. In an embodiment, step a) is carried out for 9 to 12 days, preferably for 9 to 11 days, preferably for 9 to 10 days. In an embodiment, the solution comprising growth factors and / or cytokines comprises VEGF (vascular endothelial growth factor, ThermoFisher), SCF (stem cell growth factor, ThermoFisher), FLt3-L (Fms-like tyrosine kinase 3-ligand, ThermoFisher), IL-3 (recombinant interleukin 3, ThermoFisher) and / or FGF2.

[1034] A preferred solution comprising growth factors and / or cytokines comprises VEGF (vascular endothelial growth factor, ThermoFisher), SCF (stem cell growth factor, ThermoFisher), FLt3-L (Fms-like tyrosine kinase 3-ligand, ThermoFisher), IL-3 (recombinant interleukin 3 ThermoFisher) and FGF2. Preferably, the solution comprising growth factors and / or cytokines comprises from about 2 ng / mL to about 200 ng / mL VEGF, preferably about 20 ng / mL. Preferably, the solution comprising growth factors and / or cytokines comprises from about 10 ng / mL to about 300 ng / mL of SCF, preferably about 100 ng / mL of SCF. Preferably, the solution comprising growth factors and / or cytokines comprises from about 2 ng / mL to about 200 ng / mL of Flt3L, preferably about 20 ng / mL of Flt3L. Preferably, a solution comprising growth factors and / or cytokines which does not comprise FGF2 is used just before the end of the phase of induction of the formation of embryoid bodies. Preferably, this solution is used from day D7 during the phase of induction of the formation of embryoid bodies. According to an embodiment of the invention, the embryoid bodies obtained comprise mesenchymal stem stem cells capable of expressing the CD34 marker. Thus, the inventors have shown that the first step of the method of the invention allows to obtain a subpopulation of mesenchymal stem cells exhibiting the CD34+ phenotype. More particularly, about 40% of the total cell population expresses CD34+ after 7 days of culture (first phase). The inventors have further demonstrated the obtaining of a CD34+CD43+ subpopulation and a CD34+CD43-subpopulation, these two populations being present in relatively equivalent proportions in the population. Surprisingly, the inventors have shown that the increase of the CD34+CD43+ subpopulation in the embryoid bodies, and the presence of the CD34+CD43- subpopulation in the embryoid bodies, make the total cell population more suitable for continuing with the cell differentiation protocol according to the invention.

[1035] In some embodiments of the invention mesenchymal stem cells are utilized for treatment of inflammatory conditions, these include autoimmune diseases, transplant rejection, arthritis, graft-versus-host-disease, bacterial infection, sepsis and inflammation The autoimmune diseases may, but be limited to, include at least one selected from the group consisting of Crohn's disease, erythema, atopic dermatitis, rheumatoid arthritis, Hashimoto's thyroiditis, pernicious anemia, Addison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism, hyperthyroidism, scleroderma, Behcet's disease, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, Meniere's syndrome, Guillain-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, psoriasis, vitiligo, systemic scleroderma, asthma and ulcerative colitis.ExampleExample 1: Generation of pMSC Using Decellularized Bone Matrix

[1036] Induced pluripotent stem cells are grown on feeder cells were switched to suspension culture in order to form embryoid bodies. hPSCs are cultured in E8 medium on Matrigel or vitronectin coated plates, and medium is changed daily. Two days before EB formation, 60-70% confluent hPSCs are passaged onto Matrigel or vitronectin coated plates / dishes with EDTA / PBS as previously described (Beers et al., 2012). The passage ratio is usually 1:3 or 1:4, in E8 media with 10 pM ROCK inhibitor (Y-27632) to increase cell survival. The next day, change media to E8 medium without ROCK inhibitor. On the day of EB formation when the cells grow to 60-80% confluence, cells are washed once and then incubated in EDTA / PBS for 3-15 minutes to dissociate colonies to cell clumps or single cells according to EB formation methods.3-5 minutes incubation - Selfaggregated EB formation. 10-15 minutes incubation - Forced aggregation by hanging drop or using AggreWell. Cell Harvest. Two methods are used to harvest the cells according to EDTA treatment time. 3-5 minutes EDTA incubation where most cells are still attached to plate surface. Aspirate EDTA / PBS, wash cell clumps off the plate by E8 / PVA (5 ml / dish) with ROCK inhibitor. This method is suitable for Self-aggregated EB formation. 10-15 minutes EDTA incubation where most cells detach from the plate surface to become single cells or small aggregates. Gently break the aggregates with PBS / EDTA (5 ml / dish) by pipetting, and then transfer the cells into 15 ml tube, neutralize with equal volume of E8 / PVA medium with ROCK inhibitor, count cell number, and spin the cells down at 1,000 RPM for 5 minutes. Finally, re-suspend the cells in E8 / PVA medium with ROCK inhibitor. This method is suitable for forced aggregation, such ashanging drop or using AggreWell (Mohr et al., 2010; Watanabe et al., 2007). To form self-aggregated EBs, suspend cell clumps into Corning low attachment dishes or poly- HEMA coated petri dishes via 1:1 passage (suspend the cells from one 10 cm dish in 5 ml E8 / PVA media with Rock inhibitor to one 10 cm low attachment dish to allow selfaggregation in 37°C incubator overnight. On the second day, EBs should form in various sizes. To form hanging drop EBs, single cell drops (2000 cells / 20 uL) are hanging cultured on the lid of Petri dishes. Incubate the dishes in 37°C incubator overnight. On the second day EBs should form with uniform size. The aggregated EBs can be washed off into Corning low attachment dishes or poly-HEMA coated petri dishes (Lin et al., 2014).

[1037] To form EBs in AggreWells, rinse each well of AggreWell-800 by DMEM / F12 before use. Add 0.5 mL E8 / PVA medium to each well that will be used and centrifuge to remove air bubbles. Add 1.5 x 106 cells / 1.5 mL into each well (5000 cells / microwell) to generate the desired size of EBs. Centrifuge the AggreWell plate at 100 x g, 3 mins to capture cells in the microwells. Incubate the plate in 37°C incubator overnight. On the second day EB should form with uniform size. Gently pipet the medium up and down in AggreWell to remove the EBs from microwells and transfer them into Corning low attached dishes or poly-HEMA coated petri dishes (Stem Cell Technologies technical manual). After EBs are formed and transferred to Corning low attachment dishes or poly-HEMA coated petri dishes, tilt the plate at 30°-45° angle to allow the EBs to gather at the bottle of well. Gently remove most medium with pipette, and change media to desired differentiation conditions for specific cell type differentiation. For example, in spontaneous differentiation, the medium is switched to E6 medium to culture 9 to 14 days. The medium can be changed every 2 days.

[1038] Embryoid bodies are subsequently dissociated used the Embyoid Body Dissociation Kit from Milteny Biotec following the manufacturers instructions. Briefly, transfer the EB-containing medium to a tube and centrifuge, Discard the supernatant and add DPBS to resuspend the EBs, Centrifuge again and discard the supernatant, Add the enzyme mix and incubate for 10 minutes, Use a pipette to mechanically dissociate the EBs by pipetting up and down for one minute.

[1039] Decellularized bone matrix was produced by taking human bones were purchased from Tissue Source, LLC (Lafayette, IN, USA). In brief, the native bone was placed on a tissue holder, which was then inserted into a SCCO2 vessel system (Helix SFE Version R3U, Applied Separations Inc., Allentown, PA, USA) containing 30 mL 95%ethanol. The SCCO2 system was then operated at 350 bar and 45 °C for 80 min to produce decellularized bone matrix.

[1040] Dissociated embryoid body cells are cultured with bone matrix and assessed for markers of mesenchymal stem cells by flow cytometry. Culture is performed in EMEM media supplemented with conditioned media from cultured mesenchymal stem cells at 5% volume by volume.

[1041] The mesenchymal stem cell associated protein CD105 was assessed by flow cytometry and expressed as mean fluorescent intensity (MFI). Results are shown in FIG.1.

Claims

Claims1. A method of generating mesenchymal stem cells comprising the steps of: a) obtaining a pluripotent stem cell; b) contacting said pluripotent stem cell with a matrix inductive of mesenchymal stem cell differentiation; c) seeding said matrix with mesenchymal stem cell stimulatory growth factors; and d) isolating said resulting mesenchymal stem cells.

2. The method of claim 1, wherein said pluripotent stem cell expresses hTERT.

3. The method of claim 1, wherein said pluripotent stem cell is an induced pluripotent stem cell.

4. The method of claim 1, wherein said matrix inductive of mesenchymal stem cell differentiation is decellularized bone.

5. The method of claim 4, wherein said decellularized bone possesses intact TGF-beta molecules.

6. The method of claim 5, wherein said decellularized bone contains FGF-1 at 10-1000 pg of FGF- 1 per gram of bone tissue.

7. The method of claim 5, wherein said decellularized bone contains BMP-2 at 100-5000 pg of BMP-2 per gram of bone tissue.

8. The method of claim 4, wherein said decellularized bone possesses intact granulocyte colony stimulating factor.

9. The method of claim 4, wherein said decellularized bone possesses intact granulocyte monocyte colony stimulating factor.

10. The method of claim 4, wherein said decellularized bone possesses intact macrophage colony stimulating factor.

11. The method of claim 4, wherein said decellularized bone possesses intact angiopoietin.

12. The method of claim 1, wherein said pluripotent stem cells are allowed to form embryoid bodies before contacting with said mesenchymal stem cell inductive matrix.

13. The method of claim 12, wherein said embryoid bodies are formed by culture of said uripotent stem cells in absence of feeder cells.

14. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of leukemia inhibitory factor.

15. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of interleukin-3.

16. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of interleukin-6.

17. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of interleukin-11.

18. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of interleukin-35.

19. The method of claim 13, wherein said culture in absence of feeder cells is performed in te presence of interleukin-37.

20. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of steel factor.

21. The method of claim 13, wherein said culture in absence of feeder cells is performed in the presence of osteopontin.

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