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.

1 . An in vitro cell culture comprising human ectoderm cells and human definitive endoderm cells, wherein said definitive endoderm cells are multipotent cells that can differentiate into cells of the gut tube or organs derived therefrom, and wherein said ectoderm cells comprise less than 5% of the cells in said cell culture. 
     
     
         2 . The cell culture of  claim 1 , wherein said ectoderm cells and said definitive endoderm cells together comprise greater than 10% of the cells in said cell culture. 
     
     
         3 . The cell culture of  claim 1 , wherein said ectoderm cells and said definitive endoderm cells together comprise greater than 15% of the cells in said cell culture. 
     
     
         4 . The cell culture of  claim 1  further comprising a human pluripotent stem cell. 
     
     
         5 . The cell culture of  claim 1  further comprising an extraembryonic endoderm cell. 
     
     
         6 . The cell culture of  claim 5  further comprising a human pluripotent stem cell. 
     
     
         7 . The cell culture of  claim 6 , wherein said human pluripotent stem cell comprises an embryonic stem cell. 
     
     
         8 . The cell culture of  claim 7 , wherein said embryonic stem cell is derived from a tissue selected from the group consisting of the morula and the inner cell mass (ICM) of an embryo. 
     
     
         9 . The cell culture of  claim 7 , wherein said human pluripotent cell is derived from a gondal ridge of an embryo. 
     
     
         10 . The cell culture of  claim 7  further comprising a medium. 
     
     
         11 . The cell culture of  claim 10 , wherein said medium lacks B27. 
     
     
         12 . The cell culture of  claim 10 , wherein said medium lacks serum replacement. 
     
     
         13 . The cell culture of  claim 10 , wherein said medium comprises a growth factor of the Nodal/Activin subgroup of the TGFβ superfamily. 
     
     
         14 . The cell culture of  claim 10 , further comprising a growth factor selected from the group consisting of Nodal, Activin A, Activin B and combinations thereof. 
     
     
         15 . The cell culture of  claim 14 , wherein said medium comprises a Wnt family member.