Assessing the function of mTOR in human embryonic stem cells

Jiaxi Zhou, Dong Li, Fei Wang

Research output: Chapter in Book/Report/Conference proceedingChapter


We described a protocol for dissecting the function of an important serine/threonine protein kinase, mammalian target of rapamycin (mTOR), in regulating the long-term undifferentiated growth of human embryonic stem cells (hESCs). The function of mTOR in hESCs was inactivated with a highly specific chemical inhibitor, rapamycin, and gene-specific small-hairpin RNAs, and the effects were evaluated under self-renewal or early differentiation conditions. We found that inactivation of mTOR impairs proliferation and enhances mesoderm and endoderm activities of hESCs. This protocol described a general strategy for studying the function of key genes and signaling events during hESC long-term self-renewal and early lineage specifications with pharmacological and genetic approaches.

Original languageEnglish (US)
Title of host publicationmTOR
Subtitle of host publicationMethods and Protocols
EditorsThomas Weichhart
Number of pages12
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Differentiation
  • Human embryonic stem cell
  • Long-term self-renewal
  • mTOR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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