Expansion of definitive endoderm cells

Abstract

Disclosed herein are cell cultures comprising expanded definitive endoderm cells as well as methods for expanding definitive endoderm cells in culture.

1 .- 11 . (canceled) 
     
     
         12 . An in vitro population of passaged human definitive endoderm cells. 
     
     
         13 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells expresses SOX17, GSC, MIXL1 or CXCR4. 
     
     
         14 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells expresses SOX17 and CXCR4. 
     
     
         15 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells does not express Oct4, Brachyury, ZIC1 or SOX1. 
     
     
         16 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is a population of cells that has been passaged at least twice. 
     
     
         17 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is in contact with a surface coated with human fibronectin. 
     
     
         18 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is in a medium comprising less than 2% (v/v) serum. 
     
     
         19 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is in a medium comprising at least one growth factor from the TGFβ superfamily or a member of the epidermal growth factor family. 
     
     
         20 . The cell population of  claim 19 , wherein the at least one growth factor from the TGFβ superfamily comprises activin A. 
     
     
         21 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells has been passaged using a protease or by disrupting contacts between cells. 
     
     
         22 . The cell population of  claim 21 , wherein the protease is trypsin. 
     
     
         23 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is derived from human embryonic stem cells (hESCs). 
     
     
         24 . The cell population of  claim 12 , wherein the population of passaged human definitive endoderm cells is substantially free of pluripotent cells. 
     
     
         25 . The cell population of  claim 24 , wherein at least 1 passaged human definitive endoderm cell is present for every 5 pluripotent cells. 
     
     
         26 . A method of making passaged human definitive endoderm cells in vitro comprising passaging definitive endoderm cells thereby forming passaged human definitive endoderm cells. 
     
     
         27 . The method of  claim 26 , wherein the passaged human definitive endoderm cells express SOX17, GSC, MIXL1 or CXCR4. 
     
     
         28 . The method of  claim 26 , wherein the passaged human definitive endoderm cells do not express Oct4, Brachyury, ZIC1 or SOX1. 
     
     
         29 . The method of  claim 26 , wherein the definitive endoderm cells are passaged at least twice. 
     
     
         30 . The method of  claim 26 , wherein the definitive endoderm cells are derived from human embryonic stem cells (hESCs). 
     
     
         31 . The method of  claim 26 , wherein 1 passaged human definitive endoderm cell is present for every 5 pluripotent cell. 
     
     
         32 . A method of expanding human definitive endoderm cells in vitro comprising passaging definitive endoderm cells thereby expanding human definitive endoderm cells.