Directed induction of anterior and posterior primitive streak by Wnt from embryonic stem cells cultured in a chemically defined serum-free medium

FASEB J. 2009 Jan;23(1):114-22. doi: 10.1096/fj.08-111203. Epub 2008 Sep 22.

Abstract

Formation of the primitive streak (PS) is the initial specification step that generates all the mesodermal and endodermal tissue lineages during early differentiation. Thus, a therapeutically compatible and efficient method for differentiation of the PS is crucial for regenerative medicine. In this study, we developed chemically defined serum-free culture conditions for the differentiation of embryonic stem (ES) cells into the PS-like cells. Cultures supplemented with Wnt showed induction of expression of a PS marker, the brachyury gene, followed by induction of the anterior PS markers goosecoid and foxa2, a posterior PS marker, evx1, and the endoderm marker sox17. Similar differentiation of PS by Wnt was also observed in human ES cells. Moreover, we revealed that the activation of the Wnt canonical pathway is essential for PS differentiation in mouse ES cells. These results demonstrated that Wnt is an essential and sufficient factor for the induction of the PS-like cells in vitro. These conditions of induction could constitute the initial step in generating therapeutically useful cells of the definitive endoderm lineage, such as hepatocytes and pancreatic endocrine cells, under chemically defined conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activins / pharmacology
  • Animals
  • Cell Culture Techniques
  • Cell Differentiation
  • Culture Media, Serum-Free
  • Dose-Response Relationship, Drug
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / drug effects*
  • Gene Expression Regulation, Developmental / physiology
  • Humans
  • Mice
  • Primitive Streak / drug effects*
  • Primitive Streak / physiology*
  • Signal Transduction
  • Wnt Proteins / administration & dosage
  • Wnt Proteins / pharmacology*
  • Wnt3 Protein

Substances

  • Culture Media, Serum-Free
  • Wnt Proteins
  • Wnt3 Protein
  • Activins