Method to Synchronize Cell Cycle of Human Pluripotent Stem Cells without Affecting Their Fundamental Characteristics.

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Author
Yiangou, L; Grandy, RA; Morell, CM; Tomaz, RA; Osnato, A; Kadiwala, J; Muraro, D; Garcia-Bernardo, J; Nakanoh, S; Bernard, WG; ...Date
2019-01-08Source Title
Stem Cell ReportsPublisher
Elsevier BVUniversity of Melbourne Author/s
McCarthy, DavisAffiliation
School of Mathematics and StatisticsMetadata
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Journal ArticleCitations
Yiangou, L., Grandy, R. A., Morell, C. M., Tomaz, R. A., Osnato, A., Kadiwala, J., Muraro, D., Garcia-Bernardo, J., Nakanoh, S., Bernard, W. G., Ortmann, D., McCarthy, D. J., Simonic, I., Sinha, S. & Vallier, L. (2019). Method to Synchronize Cell Cycle of Human Pluripotent Stem Cells without Affecting Their Fundamental Characteristics.. Stem Cell Reports, 12 (1), pp.165-179. https://doi.org/10.1016/j.stemcr.2018.11.020.Access Status
Open AccessOpen Access at PMC
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6335580Abstract
Cell cycle progression and cell fate decisions are closely linked in human pluripotent stem cells (hPSCs). However, the study of these interplays at the molecular level remains challenging due to the lack of efficient methods allowing cell cycle synchronization of large quantities of cells. Here, we screened inhibitors of cell cycle progression and identified nocodazole as the most efficient small molecule to synchronize hPSCs in the G2/M phase. Following nocodazole treatment, hPSCs remain pluripotent, retain a normal karyotype and can successfully differentiate into the three germ layers and functional cell types. Moreover, genome-wide transcriptomic analyses on single cells synchronized for their cell cycle and differentiated toward the endoderm lineage validated our findings and showed that nocodazole treatment has no effect on gene expression during the differentiation process. Thus, our synchronization method provides a robust approach to study cell cycle mechanisms in hPSCs.
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