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Stem Cell Reports. 2014 Jun 12;3(1):142-55. doi: 10.1016/j.stemcr.2014.05.006. eCollection 2014 Jul 8.

Time-lapse analysis of human embryonic stem cells reveals multiple bottlenecks restricting colony formation and their relief upon culture adaptation.

Author information

1
Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, Western Bank, Sheffield S10 2TN, UK ; Department of Materials Science and Engineering, The University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
2
Centre for Signal Processing and Complex Systems, Department of Automatic Control and Systems Engineering, The University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
3
Centre for Stem Cell Biology, Department of Biomedical Science, The University of Sheffield, Western Bank, Sheffield S10 2TN, UK.

Abstract

Using time-lapse imaging, we have identified a series of bottlenecks that restrict growth of early-passage human embryonic stem cells (hESCs) and that are relieved by karyotypically abnormal variants that are selected by prolonged culture. Only a minority of karyotypically normal cells divided after plating, and these were mainly cells in the later stages of cell cycle at the time of plating. Furthermore, the daughter cells showed a continued pattern of cell death after division, so that few formed long-term proliferating colonies. These colony-forming cells showed distinct patterns of cell movement. Increasing cell density enhanced cell movement facilitating cell:cell contact, which resulted in increased proportion of dividing cells and improved survival postplating of normal hESCs. In contrast, most of the karyotypically abnormal cells reentered the cell cycle on plating and gave rise to healthy progeny, without the need for cell:cell contacts and independent of their motility patterns.

PMID:
25068128
PMCID:
PMC4110749
DOI:
10.1016/j.stemcr.2014.05.006
[Indexed for MEDLINE]
Free PMC Article

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