title: 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.
authors: Yale Stem Cell Center,Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors: Cynthia M
authors: CM
authors: Megyola
authors: Yuan
authors: Y
authors: Gao
authors: Alexandra M
authors: AM
authors: Teixeira
authors: Jijun
authors: J
authors: Cheng
authors: Kartoosh
authors: K
authors: Heydari
authors: Ee-Chun
authors: EC
authors: Cheng
authors: Timothy
authors: T
authors: Nottoli
authors: Diane S
authors: DS
authors: Krause
authors: Jun
authors: J
authors: Lu
authors: Shangqin
authors: S
authors: Guo
mesh_terms: D000818:Animals; D015820:Cadherins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002450:Cell Communication / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology; D059008:Time-Lapse Imaging / Q000379:methods
chemical_list: D015820:Cadherins