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

Stem Cells. 2005 Sep;23(8):1035-43. doi: 10.1634/stemcells.2005-0080. Epub 2005 Jun 27.


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 hCG-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 gatekeeper of pluripotency in human embryonic stem and carcinoma cells by preventing their differentiation to extraembryonic endoderm and trophectoderm lineages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blastocyst / metabolism
  • Cell Differentiation / physiology*
  • Cell Line, Tumor
  • Cell Lineage / physiology
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / physiology*
  • Down-Regulation
  • Embryo, Mammalian / cytology
  • Embryonal Carcinoma Stem Cells
  • GATA4 Transcription Factor / metabolism
  • GATA6 Transcription Factor / metabolism
  • Gene Expression Regulation, Developmental
  • Germ Layers / cytology
  • Germ Layers / metabolism
  • Green Fluorescent Proteins
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / physiology*
  • Humans
  • Nanog Homeobox Protein
  • Neoplastic Stem Cells
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • RNA Interference
  • RNA, Messenger / metabolism
  • Transfection
  • alpha-Fetoproteins / metabolism


  • DNA-Binding Proteins
  • GATA4 Transcription Factor
  • GATA4 protein, human
  • GATA6 Transcription Factor
  • GATA6 protein, human
  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • RNA, Messenger
  • alpha-Fetoproteins
  • Green Fluorescent Proteins