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


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.

SUBMITTER: Megyola CM 

PROVIDER: S-EPMC4309553 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature

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

Megyola Cynthia M CM   Gao Yuan Y   Teixeira Alexandra M AM   Cheng Jijun J   Heydari Kartoosh K   Cheng Ee-Chun EC   Nottoli Timothy T   Krause Diane S DS   Lu Jun J   Guo Shangqin S  

Stem cells (Dayton, Ohio) 20130501 5


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  ...[more]

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