A novel chemically directed route for the generation of definitive endoderm from human embryonic stem cells based on inhibition of GSK-3

Heather K Bone, A S Nelson, C E Goldring, David Tosh, Melanie J Welham

Research output: Contribution to journalArticle

82 Citations (Scopus)

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. fetoprotein and HNF4 alpha. 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.
Original languageEnglish
Pages (from-to)1992-2000
Number of pages9
JournalJournal of Cell Science
Volume124
Issue number12
Early online date23 May 2011
DOIs
Publication statusPublished - 15 Jun 2011

Keywords

  • embryonic stem cells
  • hepatic differentiation
  • directed differentiation
  • glycogen synthase kinase 3 (GSK-3) inhibition
  • definitive endoderm

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