Perivascular stromal cells from primate pluripotent stem cells

Abstract

The invention provides methods, compositions and kits for making and using pericyte-like cells or perivascular stromal cells derived from pPS cells.

1 .- 12 . (canceled) 
     
     
         13 . A method for differentiating pluripotent stem cells into pericyte-like cells, the method comprising: 1) obtaining embryoid bodies from pluripotent stem cells; 2) seeding embryoid bodies at high density in low attachment plates; 3) culturing the embryoid bodies in suspension in the presence of appropriate factors; and 4) dissociating the embryoid bodies and transferring the dissociated cells to adherent plates for further culturing in the presence of appropriate factors. 
     
     
         14 . The method of  claim 13 , wherein the seeding density of step 2) is at least 1.5×10 6  cells/mL. 
     
     
         15 . The method of  claim 13 , wherein the seeding density of step 2) is at least 3×10 6  cells/mL. 
     
     
         16 . The method of  claim 13 , wherein the cells are cultured in the presence of a BMP. 
     
     
         17 . The method of  claim 16 , wherein the BMP is BMP-4. 
     
     
         18 . The method of  claim 13 , wherein further culturing of step 4 is performed in a medium comprising bFGF, optionally also comprising VEGF-A, optionally also comprising a TGFβ signaling inhibitor. 
     
     
         19 . The method of  claim 13 , further comprising an additional step of dissociating and harvesting of cells of step 4, and expanding said cells by further culturing on adherent plates in the presence of appropriate factors. 
     
     
         20 . The method of  claim 19 , wherein the additional step is performed in a medium not comprising BMP.