Patent ID: 8017393

Claim:
A method for generating and expanding CD34 − CD31 − human hemangio-colony forming cells in vitro, said method comprising the steps of: (a) culturing a cell culture comprising human embryo-derived 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 embryo-derived cells into embryoid bodies; (b) adding basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), bone morphogenic protein 4 (BMP-4), stem cell factor (SCF), Flt-3L (FL), thrombopoietin (TPO), and tPTD-HOXB4 to said culture comprising embryoid bodies and continuing to culture said culture in serum-free media; (c) disaggregating said embryoid bodies into single cells; and (d) culturing a cell culture comprising said single cells in serum-free media in the presence 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), and tPTD-HOXB4 in an amount sufficient to expand CD34 − CD31 − human hemangio-colony forming cells in said culture comprising said single cells, wherein said embryo-derived cells, embryoid bodies and CD34 − CD31 − hemangio-colony forming cells are grown in serum-free media throughout steps (a)-(d) of said method.