Two-Phase Analysis of Molecular Pathways Underlying Induced Pluripotent Stem Cell Induction§

Authors

  • Zhaoyu Lin,

    1. MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, People's Republic of China
    2. Fay Simon Center for Hearing Research at the Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri, USA
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  • Philip Perez,

    1. Fay Simon Center for Hearing Research at the Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri, USA
    2. Division of Biology & Biomedical Science and Neuroscience Program, Washington University School of Medicine, St. Louis, Missouri, USA
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  • Debin Lei,

    1. Fay Simon Center for Hearing Research at the Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri, USA
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  • Jingyue Xu,

    1. MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, People's Republic of China
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  • Xiang Gao,

    1. MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center of Nanjing University, Nanjing, Jiangsu, People's Republic of China
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  • Jianxin Bao

    Corresponding author
    1. Fay Simon Center for Hearing Research at the Department of Otolaryngology, Washington University School of Medicine, St. Louis, Missouri, USA
    2. Division of Biology & Biomedical Science and Neuroscience Program, Washington University School of Medicine, St. Louis, Missouri, USA
    • CID at Washington University School of Medicine, 4560 Clayton Avenue, St. Louis, Missouri 63110, USA
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    • Telephone: 314-747-7199; Fax: 314-747-7230


  • Author Contributions: Z.L.: conception and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; P.P.: collection and assembly of data and manuscript writing; D.L. and J.X.: collection and assembly of data; X.G.: conception and design and financial support, J.B.: conception and design, financial support, administrative support, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript.

  • Disclosure of potential conflicts of interest is found at the end of this article.

  • §

    First published online in STEM CELLSEXPRESS September 28, 2011.

Abstract

Induced pluripotent stem cells (iPSCs) can be reprogrammed from adult somatic cells by transduction with Oct4, Sox2, Klf4, and c-Myc, but the molecular cascades initiated by these factors remain poorly understood. Impeding their elucidation is the stochastic nature of the iPS induction process, which results in heterogeneous cell populations. Here we have synchronized the reprogramming process by a two-phase induction: an initial stable intermediate phase following transduction with Oct4, Klf4, and c-Myc, and a final iPS phase following overexpression of Sox2. This approach has enabled us to examine temporal gene expression profiles, permitting the identification of Sox2 downstream genes critical for induction. Furthermore, we have validated the feasibility of our new approach by using it to confirm that downregulation of transforming growth factor β signaling by Sox2 proves essential to the reprogramming process. Thus, we present a novel means for dissecting the details underlying the induction of iPSCs, an approach with significant utility in this arena and the potential for wide-ranging implications in the study of other reprogramming mechanisms. STEM Cells 2011;29:1963–1974.

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