Generating vasculogenic cell populations
The present invention relates generally to methods and compositions useful for therapeutic vascular tissue engineering. In particular, the present invention provides methods for generating substantially pure populations of vasculogenic cells from human mesenchymal progenitors, and methods and compositions for clinical applications in the field of regenerative medicine.
1. A method for generating an isolated population of primate vasculogenic cells, the method comprising:
culturing a primate mesenchymoangioblast (MAB)-derived colony comprising PDGFRβ + (Platelet Derived Growth Factor Receptor β + )/EMCN high /CD105 low /CD248 − /CD73 − /CD31 − mesenchymal progenitors on a culture plate having a coating comprising fibronectin, gelatin or collagen, in a serum-free culture medium comprising basic fibroblast growth factor (FGF2) and Platelet-Derived Growth Factor BB (PDGF-BB) in an amount effective to promote differentiation of the mesenchymal progenitors to obtain vasculogenic immature pericytes having a RGS5 + (Regulator of G-protein Signaling 5 + )/PDGFRβ + /ANG-1 + (angiopoietin-1 + )/CD 146 + /CD44 + /CD 13 + /NG2 + (Nerve/Glial antigen 2 + )/CD73 + expression profile; and
(b) isolating the vasculogenic immature pericytes.
2. The method of claim 1 , wherein the primate is human.
3. The method of claim 1 , wherein the vasculogenic immature pericytes do not express or have low expression relative to a smooth muscle cell of at least one molecular marker of smooth muscle cells selected from a-SMA, calponin, desmin, SM22, MYOCD, and MYH11.
4. The method of claim 1 , wherein the method further comprises culturing the vasculogenic immature pericytes in the presence of a maturation culture medium comprising a TGFP receptor inhibitor and PDGF-BB under conditions that promote maturation of immature pericytes to mature pericytes.
5. The method of claim 4 , wherein the TGFβ receptor inhibitor is SB-431542.
6. The method of claim 4 , wherein the mature pericyte is a NG2 high /α-SMA low /RGS5 + capillary pericyte.
7. The method of claim 4 , wherein the maturation culture medium further comprises PDGF-BB and Vascular Endothelial Growth Factor-A (VEGF), and wherein the mature pericyte is a NG2 low /α-SMA high /RGS5 + venule pericyte.
8. The method of claim 4 , wherein the maturation culture medium further comprises PDGF-BB, VEGF, and Epidermal Growth Factor 2 (EGF2), and wherein the mature pericyte is a NG2 high /α-SMA high /RGS5 + arteriole pericyte.
9. The method of claim 1 , wherein the colony of mesenchymal progenitors is clonal.
10. The method of claim 1 , wherein the colony of mesenchymal progenitors is polyclonal.