Dynamic Migration and Cell-Cell Interactions of Early Reprogramming Revealed by High-Resolution Time-Lapse Imaging§

Authors

  • Cynthia M. Megyola,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Genetics,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Yuan Gao,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Laboratory Medicine,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Alexandra M. Teixeira,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Pathology,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Jijun Cheng,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Genetics,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Kartoosh Heydari,

    1. Department of Immunobiology, Yale Flow Cytometry Core Facility,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Ee-Chun Cheng,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Cell Biology, and Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Timothy Nottoli,

    1. Department of Comparative Medicine; Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Diane S. Krause,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Laboratory Medicine,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Jun Lu,

    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Genetics,Yale University School of Medicine, New Haven, Connecticut, USA
    Search for more papers by this author
  • Shangqin Guo

    Corresponding author
    1. Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut, USA
    2. Department of Genetics,Yale University School of Medicine, New Haven, Connecticut, USA
    • 214K, 10 Amistad St., Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut 06520, USA
    Search for more papers by this author
    • Author contributions: C.M.M.: collection and/or assembly of data, data analysis and interpretation, and manuscript writing, Y.G. and T. N.: collection and/or assembly of data and data analysis and interpretation; A.M.T.: provision of study material or patients and collection and/or assembly of data; J.C. and K.H.: collection and/or assembly of data; E.C.: provision of study material or patients; D.S.K.: provision of study material or patients and manuscript writing; J.L.: conception and design, financial support, data analysis and interpretation, and manuscript writing; S.G.: conception and design, financial support, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript.

    • Telephone: 203-737-4652; Fax: 203-785-4305


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

  • §

    First published online in STEM CELLSEXPRESS January 17, 2013.

Abstract

Discovery of the cellular and molecular mechanisms of induced pluripotency has been hampered by its low efficiency and slow kinetics. Here, we report an experimental system with multicolor time-lapse microscopy that permits direct observation of pluripotency induction at single cell resolution, with temporal intervals as short as 5 minutes. Using granulocyte-monocyte progenitors as source cells, we visualized nascent pluripotent cells that emerge from a hematopoietic state. We engineered a suite of image processing and analysis software to annotate the behaviors of the reprogramming cells, which revealed the highly dynamic cell-cell interactions associated with early reprogramming. We observed frequent cell migration, which can lead to sister colonies, satellite colonies, and colonies of mixed genetic makeup. In addition, we discovered a previously unknown morphologically distinct two-cell intermediate of reprogramming, which occurs prior to other reprogramming landmarks. By directly visualizing the reprogramming process with E-cadherin inhibition, we demonstrate that E-cadherin is required for proper cellular interactions from an early stage of reprogramming, including the two-cell intermediate. The detailed cell-cell interactions revealed by this imaging platform shed light on previously unappreciated early reprogramming dynamics. This experimental system could serve as a powerful tool to dissect the complex mechanisms of early reprogramming by focusing on the relevant but rare cells with superb temporal and spatial resolution. STEM CELLS 2013;31:895–905

Ancillary