title: A novel chemically directed route for the generation of definitive endoderm from human embryonic stem cells based on inhibition of GSK-3.
abstract: The use of small molecules to 'chemically direct' differentiation represents a powerful approach to promote specification of embryonic stem cells (ESCs) towards particular functional cell types for use in regenerative medicine and pharmaceutical applications. Here, we demonstrate a novel route for chemically directed differentiation of human ESCs (hESCs) into definitive endoderm (DE) exploiting a selective small-molecule inhibitor of glycogen synthase kinase 3 (GSK-3). This GSK-3 inhibitor, termed 1m, when used as the only supplement to a chemically defined feeder-free culture system, effectively promoted differentiation of ESC lines towards primitive streak (PS), mesoderm and DE. This contrasts with the role of GSK-3 in murine ESCs, where GSK-3 inhibition promotes pluripotency. Interestingly, 1m-mediated induction of differentiation involved transient NODAL expression and Nodal signalling. Prolonged treatment of hESCs with 1m resulted in the generation of a population of cells displaying hepatoblast characteristics, that is expressing α-fetoprotein and HNF4α. Furthermore, 1m-induced DE had the capacity to mature and generate hepatocyte-like cells capable of producing albumin. These findings describe, for the first time, the utility of GSK-3 inhibition, in a chemically directed approach, to a method of DE generation that is robust, potentially scalable and applicable to different hESC lines.
authors: Centre for Regenerative Medicine and Department of Pharmacy and Pharmacology, University of Bath, Bath BA27AY, UK.
authors: Heather K
authors: HK
authors: Bone
authors: Adam S
authors: AS
authors: Nelson
authors: Christopher E
authors: CE
authors: Goldring
authors: David
authors: D
authors: Tosh
authors: Melanie J
authors: MJ
authors: Welham
references: Nat Biotechnol. 2006 Nov;24(11):1392-401
references: 17053790
references: Blood. 2008 Feb 15;111(4):1876-84
references: 18032708
references: Cell Stem Cell. 2009 Apr 3;4(4):348-58
references: 19341624
references: Biochem Biophys Res Commun. 2006 Jun 30;345(2):789-95
references: 16707109
references: Nat Biotechnol. 2005 Dec;23(12):1534-41
references: 16258519
references: Development. 2004 Dec;131(23):5817-24
references: 15525667
references: J Cell Biol. 2000 Feb 7;148(3):567-78
references: 10662781
references: Nat Rev Genet. 2007 May;8(5):368-81
references: 17387317
references: Development. 2004 Jun;131(12):2803-15
references: 15142971
references: Genes Dev. 2001 Aug 1;15(15):1998-2009
references: 11485993
references: FASEB J. 2009 Jan;23(1):114-22
references: 18809738
references: Nature. 1990 Feb 15;343(6259):657-9
references: 1689462
references: Nature. 2008 May 15;453(7193):338-44
references: 18480815
references: Development. 2006 Oct;133(19):3787-96
references: 16943279
references: Proc Natl Acad Sci U S A. 2006 Nov 7;103(45):16806-11
references: 17077151
references: Cell Stem Cell. 2008 Oct 9;3(4):402-15
references: 18940732
references: Proc Natl Acad Sci U S A. 2008 Aug 26;105(34):12301-6
references: 18719101
references: Science. 1999 Jun 18;284(5422):1998-2003
references: 10373120
references: Nat Biotechnol. 2005 Dec;23(12):1542-50
references: 16311587
references: Chem Biol. 2009 Jan 30;16(1):15-27
references: 19171302
references: N Engl J Med. 1998 May 14;338(20):1422-6
references: 9580649
references: Dev Biol. 1997 Mar 15;183(2):234-42
references: 9126297
references: J Cell Sci. 2007 Jun 15;120(Pt 12):2078-90
references: 17535850
references: Nat Rev Genet. 2008 May;9(5):329-40
references: 18398419
references: Dev Biol. 1999 Sep 15;213(2):442-56
references: 10479460
references: Hepatology. 2010 May;51(5):1754-65
references: 20301097
references: Development. 1994 Jul;120(7):1919-28
references: 7924997
references: Development. 2007 Mar;134(6):1023-34
references: 17287255
references: Nat Chem Biol. 2009 Apr;5(4):258-65
references: 19287398
references: Development. 2004 Apr;131(7):1651-62
references: 14998924
references: J Biol Chem. 1994 Aug 19;269(33):21165-76
references: 8063737
references: Nature. 1996 May 9;381(6578):155-8
references: 8610012
references: Biochem Biophys Res Commun. 2006 Apr 28;343(1):159-66
references: 16530170
references: Nat Med. 2004 Jan;10(1):55-63
references: 14702635
references: Development. 2001 May;128(10):1831-43
references: 11311163
references: Genes Dev. 2003 Jul 1;17(13):1646-62
references: 12842913
references: Development. 2005 Oct;132(19):4363-74
references: 16141227
references: Dev Biol. 2006 Feb 1;290(1):81-91
references: 16330017
references: J Biol Chem. 2004 Oct 22;279(43):45076-84
references: 15308665
references: Nat Biotechnol. 2006 Nov;24(11):1402-11
references: 17086172
references: Nat Genet. 2003 Jul;34(3):292-6
references: 12808453
references: J Biotechnol. 2010 Feb 1;145(3):284-94
references: 19932139
references: Stem Cells. 2008 May;26(5):1117-27
references: 18292207
references: Stem Cells. 2007 Jan;25(1):29-38
references: 17204604
references: Stem Cells. 2005 Nov-Dec;23(10):1489-501
references: 16002782
references: Nat Genet. 1999 Aug;22(4):361-5
references: 10431240
references: Mech Dev. 1999 Sep;87(1-2):189-92
references: 10495285
references: Curr Drug Metab. 2006 Aug;7(6):629-60
references: 16918317
references: Development. 2008 Sep;135(17):2969-79
references: 18667462
references: Nature. 2001 Jun 21;411(6840):965-9
references: 11418863