Dynamic migration and cell-cell interactions of early reprogramming revealed by high-resolution time-lapse imaging

Stem Cells. 2013 May;31(5):895-905. doi: 10.1002/stem.1323.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cadherins / antagonists & inhibitors
  • Cadherins / metabolism
  • Cell Communication / physiology*
  • Cell Movement / physiology*
  • Cells, Cultured
  • Cellular Reprogramming / physiology*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / physiology
  • Female
  • Mice
  • Mice, Inbred C57BL
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / physiology
  • Time-Lapse Imaging / methods

Substances

  • Cadherins