HEMANGIO-COLONY FORMING CELLS

1 - 232 . (canceled)

233 . A method, comprising:

(a) culturing human embryonic stem cells in serum free media in the presence of vascular endothelial growth factor (VEGF) and bone morphogenic protein 4 (BMP-4) in an amount sufficient to induce the differentiation of said embryonic stem cells into embryoid bodies; and

(b) adding at least one first growth factor to said culture comprising the embryoid bodies to generate CD34 − CD31 − human hemangio-colony forming cells,

wherein the embryonic stem cells, embryoid bodies, and CD34 − CD31 − hemangio-colony forming cells are continuously grown in serum-free media to generate CD34 − CD31 − human hemangio-colony forming cells.

234 . The method of claim 233 , wherein the VEGF and BMP-4 are added to step (a) within 0-48 hours of cell culture, and optionally, multiple times throughout step (a).

235 . The method of claim 233 , wherein the at least one first growth factor is selected from the group consisting of: basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), bone morphogenic protein 4 (BMP-4), stem cell factor (SCF), thrombopoietin (TPO), Flt-3L (FL), HOX protein, erythropoietin (EPO) and combinations thereof.

236 . The method of claim 233 , wherein the at least one first growth factor is added to step (b) within 48-72 hours of cell culture, and optionally, added multiple times throughout step (b).

237 . The method of claim 235 , wherein the HOX protein is HOXB4, a functional equivalent or an active fragment thereof, and the functional equivalent or an active fragment thereof possess the same or substantially similar properties as wild-type HOXB4.

238 . The method of claim 233 , further comprising:

(c) disaggregating the embryoid bodies into single cells; and

(d) adding at least one second growth factor to said culture in an amount sufficient to expand CD34 − CD31 − human hemangio-colony forming cells,

wherein the embryonic stem cells, embryoid bodies, and CD34 − CD31 − hemangio-colony forming cells are continuously grown in serum-free media to expand CD34 − CD31 − human hemangio-colony forming cells.

239 . The method of claim 238 , wherein the at least one second growth factor is selected from the group consisting of: insulin, transferrin, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), erythropoietin (EPO), stem cell factor (SCF), vascular endothelial growth factor (VEGF), bone morphogenic protein 4 (BMP-4), HOX protein, thrombopoietin (TPO), Flt-3L (FL), and combinations thereof.

240 . The method of claim 238 , wherein the at least one second growth factor is added to step (d) after 72 hours of cell culture and optionally, added multiple times throughout step (d).

241 . The method of claim 239 , wherein the HOX protein is HOXB4, a functional equivalent or an active fragment thereof, and the functional equivalent or an active fragment thereof possess the same or substantially similar properties as wild-type HOXB4.

242 . The method of claim 241 , wherein the functional equivalent or an active fragment thereof, comprises a fusion protein with a protein transduction domain (PTD).

243 . The method of claim 242 , wherein the fusion protein comprises one or more copies of SEQ ID NO:14.

244 . The method of claim 238 , further comprising purifying the CD34 − CD31 − human hemangio-colony forming cells.

245 . The method of claim 244 , further comprising the use of an anti-CD71 antibody to purify the CD34 − CD31 − human hemangio-colony forming cells.

246 . The method of claim 238 , further comprising isolating the CD34 − CD31 − human hemangio-colony forming cells.

247 . The method of claim 233 , wherein the embryonic stem cells are from a library of embryonic stem cells that are hemizygous or homozygous for at least one majorhistocompatibility (MHC) allele present in a human population, and each member of the library is hemizygous or homozygous for a unique set of MHC alleles compared to other members of the library of embryonic stem cells, thereby generating a library of CD34 − CD31 − human hemangio-colony forming cells that are hemizygous or homozygous for at least one majorhistocompatibility (MHC) allele present in a human population, and each member of the library of CD34 − CD31 − human hemangio-colony forming cells is hemizygous or homozygous for a unique set of MHC alleles compared to other members of the library of human hemangio-colony forming cells.

248 . The method of claim 247 , wherein the embryonic stem cells are hemizygous or homozygous all majorhistocompatibility (MHC) alleles present in a human population, thereby generating a library of CD34 − CD31 − human hemangio-colony forming cells that are hemizygous or homozygous all majorhistocompatibility (MHC) alleles present in a human population.

249 . The method of claim 238 , further comprising differentiating CD34 − CD31 − human hemangio-colony forming cells into human hematopoietic cells.

250 . The method of claim 238 , further comprising differentiating CD34 − CD31 − human hemangio-colony forming cells into human endothelial cells.

251 . A method of administering to a patient in need thereof, the human hematopoietic cells of claim 249 , comprising:

selecting the patient;

providing a quantity of hematopoietic cells; and

administering the quantity of hematopoietic cells to the patient.

252 . The method of claim 251 , wherein the patient is in need of a blood transfusion.

253 . A method of administering to a patient in need thereof, the human endothelial cells of claim 250 , comprising:

selecting the patient;

providing a quantity of endothelial cells; and

administering the quantity of endothelial cells to the patient.

254 . A method of expanding CD34 − CD31 − human hemangio-colony forming cells, comprising:

providing a quantity of CD34 − CD31 − human hemangio-colony forming cells; and

adding at least one growth factor in an amount sufficient to expand CD34 − CD31 − human hemangio-colony forming cells, wherein the CD34 − CD31 − hemangio-colony forming cells are continuously grown in serum-free media.

255 . The method of claim 254 , wherein the at least one growth factor is selected from the group consisting of: insulin, transferrin, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), erythropoietin (EPO), stem cell factor (SCF), vascular endothelial growth factor (VEGF), bone morphogenic protein 4 (BMP-4), HOX protein, thrombopoietin (TPO), Flt-3L (FL), and combinations thereof.

256 . The method of claim 254 , further comprising differentiating CD34 − CD31 − human hemangio-colony forming cells into human hematopoietic cells cells.

257 . The method of claim 254 , further comprising differentiating CD34 − CD31 − human hemangio-colony forming cells into human endothelial cells.

258 . A method of administering to a patient in need thereof, the human hematopoietic cells of claim 256 , comprising:

selecting the patient;

providing a quantity of hematopoietic cells; and

administering the quantity of hematopoietic cells to the patient.

259 . The method of claim 258 , wherein the patient is in need of a blood transfusion.

260 . A method of administering to a patient in need thereof, the human endothelial cells of claim 257 , comprising:

selecting the patient;

providing a quantity of endothelial cells; and

administering the quantity of endothelial cells to the patient.

261 . A method of inducing tolerance in a human subject receiving a donor allograft, comprising:

(a) creating thymic space in the human subject;

(b) depleting or inactivating donor-reactive T cells in the human subject;

(c) introducing into the human subject, the human hemangio-colony forming cells generated by the method of claim 1 , wherein said hemangio-colony forming cells are match with respect to the donor, and

(d) implanting the allograft into the human subject, wherein said human hemangio-colony forming cells induce tolerance in the human subject to the allograft.