Definitive endoderm

Abstract

Disclosed herein are cell cultures comprising definitive endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified definitive endoderm cells as well as methods for enriching, isolating and purifying definitive endoderm cells from other cell types.

What is claimed is: 
     
         1 . An in vitro method of making a cell population comprising definitive endoderm cells, the method comprising:
 (a) obtaining a cell population comprising human pluripotent stem cells in medium comprising no serum or serum replacement; and   (b) providing the cell population comprising human pluripotent stem cells with a TGFβ super family member,   
       thereby producing a cell population comprising definitive endoderm cells. 
     
     
         2 . The method of  claim 1 , wherein the TGFβ super family member is activin A. 
     
     
         3 . The method of  claim 2 , comprising providing the cell population comprising human pluripotent stem cells with about 10-100 ng/ml activin A. 
     
     
         4 . The method of  claim 1 , wherein the cell population comprising definitive endoderm further comprises mesendoderm. 
     
     
         5 . The method of  claim 1 , wherein the method further comprises providing a medium comprising less than about 10% volume/volume (v/v) serum about 24 hours after the TGFβ super family member is provided. 
     
     
         6 . The method of  claim 1 , wherein the method further comprises providing a medium comprising less than about 2% volume/volume (v/v) serum about 24 hours after the TGFβ super family member is provided. 
     
     
         7 . The method of  claim 1 , wherein the method further comprises providing a medium comprising less than about 2% volume/volume (v/v) serum about 24 hours after the TGFβ super family member is provided. 
     
     
         8 . The method of  claim 1 , further comprising providing the cell population comprising human pluripotent stem cells with a WNT protein. 
     
     
         9 . The method of  claim 8 , wherein the WNT protein in Wnt3A. 
     
     
         10 . The method of  claim 1 , wherein the cell population comprising definitive endoderm cells is substantially free of cells derived from the ectoderm lineage. 
     
     
         11 . An in vitro method of making a cell population comprising definitive endoderm cells comprising:
 (a) obtaining a cell population comprising human pluripotent stem cells in a medium comprising less than about 10% serum; and   (b) providing the cell population with a TGFβ super family member,   thereby producing a cell population comprising definitive endoderm cells.   
     
     
         12 . The method of  claim 11 , wherein the TGFβ super family member is activin A. 
     
     
         13 . The method of  claim 12 , comprising providing the cell population comprising human pluripotent stem cells with about 10-100 ng/ml activin A. 
     
     
         14 . The method of  claim 11 , wherein the cell population comprising definitive endoderm further comprises mesendoderm. 
     
     
         15 . The method of  claim 11 , wherein the media comprises less than about 2% (v/v) serum. 
     
     
         16 . The method of  claim 11 , wherein the media comprises less than about 0.5% (v/v) serum. 
     
     
         17 . The method of  claim 11 , further comprising providing the cell population comprising human pluripotent stem cells with a WNT family member. 
     
     
         18 . The method of  claim 11 , wherein the WNT family member in Wnt3A. 
     
     
         19 . The method of  claim 11 , wherein the cell population comprising definitive endoderm cells is substantially free of cells derived from the ectoderm lineage. 
     
     
         20 . A method of differentiating human pluripotent cells, said method comprising culturing a population of pluripotent human cells in a medium comprising no serum or serum replacement and a TGFβ superfamily growth factor thereby differentiating the human pluripotent cells.