Robust self-renewal of rat embryonic stem cells requires fine-tuning of glycogen synthase kinase-3 inhibition

Stem Cell Reports. 2013 Aug 22;1(3):209-17. doi: 10.1016/j.stemcr.2013.07.003. eCollection 2013.


Germline-competent embryonic stem cells (ESCs) have been derived from mice and rats using culture conditions that include an inhibitor of glycogen synthase kinase 3 (GSK3). However, rat ESCs remain susceptible to sporadic differentiation. Here, we show that unsolicited differentiation is attributable to overinhibition of GSK3. The self-renewal effect of inhibiting GSK3 is mediated via β-catenin, which abrogates the repressive action of TCF3 on core pluripotency genes. In rat ESCs, however, GSK3 inhibition also leads to activation of differentiation-associated genes, notably lineage specification factors Cdx2 and T. Lowered GSK3 inhibition reduces differentiation and enhances clonogenicity and self-renewal. The differential sensitivity of rat ESCs to GSK3 inhibition is linked to elevated expression of the canonical Wnt pathway effector LEF1. These findings reveal that optimal GSK3 inhibition for ESC propagation is influenced by the balance of TCF/LEF factors and can vary between species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CDX2 Transcription Factor
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism*
  • Germ Cells / cytology
  • Germ Cells / metabolism*
  • Glycogen Synthase Kinase 3 / antagonists & inhibitors
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 beta
  • Homeodomain Proteins / metabolism
  • Lymphoid Enhancer-Binding Factor 1 / metabolism
  • Mice
  • Rats
  • Transcription Factors / metabolism
  • Wnt Signaling Pathway / genetics*
  • beta Catenin / genetics


  • CDX2 Transcription Factor
  • Cdx2 protein, rat
  • Homeodomain Proteins
  • Lef1 protein, rat
  • Lymphoid Enhancer-Binding Factor 1
  • Transcription Factors
  • beta Catenin
  • Glycogen Synthase Kinase 3 beta
  • Glycogen Synthase Kinase 3