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An optimized small molecule inhibitor cocktail supports long-term maintenance of human embryonic stem cells

Author

Listed:
  • Hideaki Tsutsui

    (University of California at Los Angeles School of Engineering and Applied Science)

  • Bahram Valamehr

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
    Present addresses: Fate Therapeutics Inc. 3535 General Atomics Court Suite 200, San Diego, California 92121, USA (B.V.); ISIS Pharmaceuticals, 1896 Rutherford Road, Carlsbad, California 92008, USA (S.G.).)

  • Antreas Hindoyan

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.)

  • Rong Qiao

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
    University of California at Los Angeles School of Medicine)

  • Xianting Ding

    (University of California at Los Angeles School of Engineering and Applied Science)

  • Shuling Guo

    (University of California at Los Angeles School of Medicine
    Present addresses: Fate Therapeutics Inc. 3535 General Atomics Court Suite 200, San Diego, California 92121, USA (B.V.); ISIS Pharmaceuticals, 1896 Rutherford Road, Carlsbad, California 92008, USA (S.G.).)

  • Owen N. Witte

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
    University of California at Los Angeles School of Medicine
    Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California at Los Angeles School of Medicine
    Howard Hughes Medical Institute, University of California at Los Angeles School of Medicine)

  • Xin Liu

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
    University of California at Los Angeles School of Medicine)

  • Chih-Ming Ho

    (University of California at Los Angeles School of Engineering and Applied Science)

  • Hong Wu

    (University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
    Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California at Los Angeles School of Medicine
    Institute for Molecular Medicine, University of California at Los Angeles School of Medicine)

Abstract

A major challenge in stem cell-mediated regenerative medicine is the development of defined culture systems for the maintenance of clinical-grade human embryonic stem (hES) cells. Here, we identify, using a feedback system control scheme, a unique combination of three small molecule inhibitors that enables the maintenance of hES cells on a fibronectin-coated surface through single cell passaging. After 20 passages, the undifferentiated state of the hES cells was confirmed by OCT4, SSEA4 and NANOG expressions, whereas their pluripotent potential and genetic integrity were demonstrated by teratoma formation and normal karyotype, respectively. Our study attests to the power of the feedback system control scheme to quickly pinpoint optimal conditions for desired biological activities, and provides a chemically defined, scalable and single cell passaging culture system for hES cells.

Suggested Citation

  • Hideaki Tsutsui & Bahram Valamehr & Antreas Hindoyan & Rong Qiao & Xianting Ding & Shuling Guo & Owen N. Witte & Xin Liu & Chih-Ming Ho & Hong Wu, 2011. "An optimized small molecule inhibitor cocktail supports long-term maintenance of human embryonic stem cells," Nature Communications, Nature, vol. 2(1), pages 1-8, September.
    • Handle: RePEc:nat:natcom:v:2:y:2011:i:1:d:10.1038_ncomms1165
    DOI: 10.1038/ncomms1165
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