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.
Embryonic Stem Cells/induced Pluripotent Stem Cells
Article first published online: 24 APR 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 5, pages 895–905, May 2013
How to Cite
Megyola, C. M., Gao, Y., Teixeira, A. M., Cheng, J., Heydari, K., Cheng, E.-C., Nottoli, T., Krause, D. S., Lu, J. and Guo, S. (2013), Dynamic Migration and Cell-Cell Interactions of Early Reprogramming Revealed by High-Resolution Time-Lapse Imaging. STEM CELLS, 31: 895–905. doi: 10.1002/stem.1323
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS January 17, 2013.
- Issue published online: 24 APR 2013
- Article first published online: 24 APR 2013
- Accepted manuscript online: 17 JAN 2013 10:46PM EST
- Manuscript Accepted: 20 DEC 2012
- Manuscript Received: 13 JUL 2012
- NIH. Grant Numbers: DK082982, DK086267
- Yale Center of Excellence in Molecular Hematology and Connecticut Innovations
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