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

Hideaki Tsutsui, Bahram Valamehr, Antreas Hindoyan, Rong Qiao, Xianting Ding, Shuling Guo, Owen N. Witte, Xin Liu, Chih-Ming Ho () and Hong Wu ()
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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.
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.
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
Owen N. Witte: University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
Xin Liu: University of California at Los Angeles School of Medicine, 650 CE Young Drive South, Los Angeles, California 90095-1735, USA.
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.

Nature Communications, 2011, vol. 2, issue 1, 1-8

Abstract: 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.

Date: 2011
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DOI: 10.1038/ncomms1165

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