Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor

Nat Med. 2004 Jan;10(1):55-63. doi: 10.1038/nm979. Epub 2003 Dec 21.

Abstract

Human and mouse embryonic stem cells (HESCs and MESCs, respectively) self-renew indefinitely while maintaining the ability to generate all three germ-layer derivatives. Despite the importance of ESCs in developmental biology and their potential impact on tissue replacement therapy, the molecular mechanism underlying ESC self-renewal is poorly understood. Here we show that activation of the canonical Wnt pathway is sufficient to maintain self-renewal of both HESCs and MESCs. Although Stat-3 signaling is involved in MESC self-renewal, stimulation of this pathway does not support self-renewal of HESCs. Instead we find that Wnt pathway activation by 6-bromoindirubin-3'-oxime (BIO), a specific pharmacological inhibitor of glycogen synthase kinase-3 (GSK-3), maintains the undifferentiated phenotype in both types of ESCs and sustains expression of the pluripotent state-specific transcription factors Oct-3/4, Rex-1 and Nanog. Wnt signaling is endogenously activated in undifferentiated MESCs and is downregulated upon differentiation. In addition, BIO-mediated Wnt activation is functionally reversible, as withdrawal of the compound leads to normal multidifferentiation programs in both HESCs and MESCs. These results suggest that the use of GSK-3-specific inhibitors such as BIO may have practical applications in regenerative medicine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / physiology
  • Embryo, Mammalian / cytology*
  • Enzyme Inhibitors / pharmacology*
  • Fluorescent Antibody Technique
  • Gene Expression Regulation / physiology
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors*
  • Humans
  • Indoles / pharmacology*
  • Mice
  • Oximes / pharmacology*
  • Pluripotent Stem Cells / cytology*
  • Proto-Oncogene Proteins / drug effects*
  • Proto-Oncogene Proteins / physiology
  • Wnt Proteins
  • Zebrafish Proteins*

Substances

  • 6-bromoindirubin-3'-oxime
  • Enzyme Inhibitors
  • Indoles
  • Oximes
  • Proto-Oncogene Proteins
  • Wnt Proteins
  • Zebrafish Proteins
  • Glycogen Synthase Kinase 3