Derivation conditions impact x-inactivation status in female human induced pluripotent stem cells

Kiichiro Tomoda, Kazutoshi Takahashi, Karen Leung, Aki Okada, Megumi Narita, N. Alice Yamada, Kirsten E. Eilertson, Peter Tsang, Shiro Baba, Mark P. White, Salma Sami, Deepak Srivastava, Bruce R. Conklin, Barbara Panning, Shinya Yamanaka

Research output: Contribution to journalArticlepeer-review

93 Scopus citations

Abstract

Female human induced pluripotent stem cell (hiPSC) lines exhibit variability in X-inactivation status. The majority of hiPSC lines maintain one transcriptionally active X (Xa) and one inactive X (Xi) chromosome from donor cells. However, at low frequency, hiPSC lines with two Xas are produced, suggesting that epigenetic alterations of the Xi occur sporadically during reprogramming. We show here that X-inactivation status in female hiPSC lines depends on derivation conditions. hiPSC lines generated by the Kyoto method (retroviral or episomal reprogramming), which uses leukemia inhibitory factor (LIF)-expressing SNL feeders, frequently had two Xas. Early passage Xa/Xi hiPSC lines generated on non-SNL feeders were converted into Xa/Xa hiPSC lines after several passages on SNL feeders, and supplementation with recombinant LIF caused reactivation of some of X-linked genes. Thus, feeders are a significant factor affecting X-inactivation status. The efficient production of Xa/Xa hiPSC lines provides unprecedented opportunities to understand human X-reactivation and -inactivation.

Original languageEnglish (US)
Pages (from-to)91-99
Number of pages9
JournalCell Stem Cell
Volume11
Issue number1
DOIs
StatePublished - Jul 6 2012

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Genetics
  • Cell Biology

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