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 cell culture comprising human definitive endoderm cells in a medium comprising serum, wherein less than about 5% (v/v) of the medium is serum. 
     
     
         2 . The cell culture of  claim 1 , wherein the medium further comprises a TGFβ superfamily growth factor. 
     
     
         3 . The cell culture of  claim 1 , wherein the medium further comprises a TGFβ superfamily growth factor and a Wnt family member. 
     
     
         4 . The cell culture of  claim 1 , wherein less than about 2% (v/v) of the medium is serum. 
     
     
         5 . The cell culture of  claim 1 , wherein less than about 0.2% (v/v) of the medium is serum. 
     
     
         6 . The cell culture of  claim 1 , wherein the medium does not comprise serum replacement. 
     
     
         7 . The cell culture of  claim 1 , wherein the human definitive endoderm cells are produced by a method comprising:
 (a) culturing a population of pluripotent human cells in a growth medium comprising serum and a TGFβ superfamily growth factor, thereby producing human definitive endoderm cells, wherein less than about 5% (v/v) of the growth medium is serum.   
     
     
         8 . The cell culture of  claim 7 , wherein said growth medium does not comprise serum replacement. 
     
     
         9 . The cell culture of  claim 7 , wherein said TGFβ superfamily growth factor in said growth medium is selected from the group consisting of Nodal, activin A and activin B. 
     
     
         10 . The cell culture of  claim 7 , wherein said TGFβ superfamily growth factor in said growth medium is activin A. 
     
     
         11 . The cell culture of  claim 7 , wherein at least 10 ng/ml activin A is present in said growth medium. 
     
     
         12 . The cell culture of  claim 7 , wherein at least 100 ng/ml activin A is present in said growth medium. 
     
     
         13 . The cell culture of  claim 7 , wherein said growth medium further comprises a Wnt family member. 
     
     
         14 . The cell culture of  claim 7 , wherein the Wnt family member is Wnt3a. 
     
     
         15 . The cell culture of  claim 7 , wherein the growth medium comprises less than 2% serum. 
     
     
         16 . The cell culture of  claim 7 , wherein the growth medium comprises less than 0.2% serum. 
     
     
         17 . The cell culture of  claim 7 , wherein the method comprises, prior to step (a), culturing the pluripotent human cells in a first growth medium comprising no serum for about 24 hours. 
     
     
         18 . The cell culture of  claim 17 , wherein the method comprises, prior to step (a) and after said culturing in said first growth medium, culturing the pluripotent human cells in a series of additional growth mediums comprising successively increasing concentrations of serum. 
     
     
         19 . An in vitro cell culture comprising human definitive endoderm cells in a medium comprising serum and a TGFβ superfamily growth factor, wherein less than about 5% (v/v) of the medium is serum. 
     
     
         20 . An in vitro cell culture comprising human definitive endoderm cells in a medium comprising serum, a TGFβ superfamily growth factor and a Wnt family member, wherein less than about 5% (v/v) of the medium is serum.