Methods of producing human foregut endoderm cells expressing pdx1 from human definitive endoderm

Abstract

Disclosed herein are cell cultures comprising dorsal and/or ventral PDX1-positive foregut endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified dorsal and/or ventral PDX1-positive foregut endoderm cells as well as methods for enriching, isolating and purifying dorsal and/or ventral PDX1-positive foregut endoderm cells from other cell types. Methods of identifying differentiation factors capable of promoting the differentiation of dorsal and/or ventral PDX1-positive foregut endoderm cells, are also disclosed.

What is claimed is: 
     
         1 . A method of producing human pancreatic-duodenal homeobox factor-1 (PDXI)-positive ventrally-biased foregut endoderm cells, comprising: culturing human definitive endoderm cells in a medium comprising an effective amount of a fibroblast growth factor (FGF)-family growth factor to promote differentiation of the human definitive endoderm cells to produce PDX I-positive ventrally-biased foregut endoderm cells. 
     
     
         2 . The method of  claim 1 , wherein the FGF-family growth factor is FGF-7 or FGF-10. 
     
     
         3 . The method of  claim 1 , wherein the medium comprises the FGF-family growth factor at a concentration of 5 ng/ml to 500 ng/ml. 
     
     
         4 . A method of producing human PDXI-positive dorsally-biased foregut endoderm cells, comprising: culturing human definitive endoderm cells in a medium comprising an effective amount of a retinoid to promote differentiation of the human definitive endoderm cells to produce PDX I-positive dorsally-biased foregut endoderm cells. 
     
     
         5 . The method of  claim 4 , wherein the retinoid is RA. 
     
     
         6 . The method of  claim 5 , wherein the medium comprises RA at a concentration of 0.2 μM to 2.0 μM.