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Preventing erosion of X-chromosome inactivation in human embryonic stem cells

Author

Listed:
  • Marissa Cloutier

    (University of Michigan Medical School)

  • Surinder Kumar

    (University of Michigan Medical School
    University of Michigan Medical School)

  • Emily Buttigieg

    (University of Michigan Medical School)

  • Laura Keller

    (University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School)

  • Brandon Lee

    (University of Michigan Medical School)

  • Aaron Williams

    (University of Michigan Medical School)

  • Sandra Mojica-Perez

    (University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School)

  • Indri Erliandri

    (University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School)

  • Andre Monteiro Da Rocha

    (University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School)

  • Kenneth Cadigan

    (Cellular, and Developmental Biology, University of Michigan Medical School)

  • Gary D. Smith

    (University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School
    University of Michigan Medical School)

  • Sundeep Kalantry

    (University of Michigan Medical School)

Abstract

X-chromosome inactivation is a paradigm of epigenetic transcriptional regulation. Female human embryonic stem cells (hESCs) often undergo erosion of X-inactivation upon prolonged culture. Here, we investigate the sources of X-inactivation instability by deriving new primed pluripotent hESC lines. We find that culture media composition dramatically influenced the expression of XIST lncRNA, a key regulator of X-inactivation. hESCs cultured in a defined xenofree medium stably maintained XIST RNA expression and coating, whereas hESCs cultured in the widely used mTeSR1 medium lost XIST RNA expression. We pinpointed lithium chloride in mTeSR1 as a cause of XIST RNA loss. The addition of lithium chloride or inhibitors of GSK-3 proteins that are targeted by lithium to the defined hESC culture medium impeded XIST RNA expression. GSK-3 inhibition in differentiating female mouse embryonic stem cells and epiblast stem cells also resulted in a loss of XIST RNA expression. Together, these data may reconcile observed variations in X-inactivation in hESCs and inform the faithful culture of pluripotent stem cells.

Suggested Citation

  • Marissa Cloutier & Surinder Kumar & Emily Buttigieg & Laura Keller & Brandon Lee & Aaron Williams & Sandra Mojica-Perez & Indri Erliandri & Andre Monteiro Da Rocha & Kenneth Cadigan & Gary D. Smith & , 2022. "Preventing erosion of X-chromosome inactivation in human embryonic stem cells," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
  • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-30259-x
    DOI: 10.1038/s41467-022-30259-x
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    References listed on IDEAS

    as
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