Downregulation of NANOG induces differentiation of human embryonic stem cells to extraembryonic lineages

Louise Hyslop, Miodrag Stojkovic, Lyle Armstrong, Theresia Walter, Petra Stojkovic, Stefan Przyborski, Mary Herbert, Alison Murdoch, Tom Strachan, Majlinda Lako

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330 Citations (Scopus)


The homeobox transcription factor Nanog has been proposed to play a crucial role in the maintenance of the undifferentiated state of murine embryonic stem cells. A human counterpart, NANOG, has been identified, but its function and localization have not hitherto been described. We have used a combination of RNA interference and quantitative real-time polymerase chain reaction to study NANOG in human embryonic stem and embryonic carcinoma cells. Transfection of NANOG-specific small interfering RNAs reduced levels of NANOG transcript and protein and induced activation of the extraembryonic endoderm-associated genes GATA4, GATA6, LAMININ B1, and AFP as well as upregulation of trophectoderm-associated genes CDX2, GATA2, hCG-alpha, and ACG-beta. Immunostaining of preimplantation human embryos showed that NANOG was expressed in the inner cell mass of expanded blastocysts but not in earlier-stage embryos, consistent with a role in the maintenance of pluripotency. Taken together, our findings suggest that NANOG acts as a gate-keeper of pluripotency in human embryonic stem and carcinoma cells by preventing their differentiation to extraembryonic endoderm and trophectoderm lineages.

Original languageEnglish
Pages (from-to)1035-1043
Number of pages9
JournalStem Cells
Issue number8
Publication statusPublished - Sept 2005
Externally publishedYes


  • Human embryonic stem cells
  • Pluripotency
  • Primitive endoderm
  • Small interfering RNA
  • Trophectoderm

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