Oct-4 Knockdown Induces Similar Patterns of Endoderm and Trophoblast Differentiation Markers in Human and Mouse Embryonic Stem Cells

Stem Cells. 2004;22(2):225-35. doi: 10.1634/stemcells.22-2-225.

Abstract

The transcription factor Oct-4 is a marker of pluripotency in mouse and human embryonic stem (ES) cells. Previous studies using a tetracycline-regulated Oct-4 transgene in the ZHBTc4 cell line demonstrated that downregulation of Oct-4 expression induced dedifferentiation into trophoblast, a lineage mouse ES cells do not normally generate. We found that transfection of Oct-4-specific short interfering RNA significantly reduced expression and functional activity of Oct-4 in mouse and human ES cells, enabling its role to be compared in both cell types. In mouse ES cells, Oct-4 knockdown produced a pattern of morphological differentiation and increase in expression of the trophoblast-associated transcription factor Cdx2, similar to that triggered by suppressing the Oct-4 transgene in the ZHBTc4 cell line. In addition, downregulation of Oct-4 was accompanied by increased expression of the endoderm-associated genes Gata6 and alpha-fetoprotein, and a gene trap associated with primitive liver/yolk sac differentiation. In human ES cells, Oct-4 knockdown also induced morphological differentiation coincident with the upregulation of Gata6. The induction of Cdx2 and other trophoblast-associated genes, however, was dependent on the culture conditions. These results establish the general requirement for Oct-4 in maintaining pluripotency in ES cells. Moreover, the upregulation of endoderm-associated markers in both mouse and human ES cells points to overlap between development of trophoblast and endoderm differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avian Proteins*
  • Cell Differentiation / physiology*
  • Cells, Cultured
  • DNA Primers
  • DNA-Binding Proteins / metabolism*
  • Embryo, Mammalian / cytology
  • GATA6 Transcription Factor
  • Homeodomain Proteins / metabolism
  • Humans
  • Mice
  • Octamer Transcription Factor-3
  • RNA, Small Interfering / metabolism
  • Stem Cells / cytology*
  • Transcription Factors / metabolism*
  • alpha-Fetoproteins / metabolism

Substances

  • Avian Proteins
  • DNA Primers
  • DNA-Binding Proteins
  • GATA6 Transcription Factor
  • GATA6 protein, human
  • Gata6 protein, mouse
  • Homeodomain Proteins
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • Pou5f1 protein, mouse
  • RNA, Small Interfering
  • Transcription Factors
  • alpha-Fetoproteins