Wellcome-MRC Cambridge Stem Cell Institute, Anne McLaren Laboratory, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Surgery, University of Cambridge, Cambridge CB2 0QQ, UK; Department of Medicine, Division of Cardiovascular Medicine, University of Cambridge, Cambridge CB2 0QQ, UK; Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton CB10 1SA, UK.
Wellcome-MRC Cambridge Stem Cell Institute, Anne McLaren Laboratory, University of Cambridge, Cambridge CB2 0SZ, UK; Department of Surgery, University of Cambridge, Cambridge CB2 0QQ, UK.
Stem Cell Reports. 2019 Jan 8;12(1):165-179. doi: 10.1016/j.stemcr.2018.11.020. Epub 2018 Dec 27.
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.
细胞周期进程和细胞命运决定在人类多能干细胞（hPSCs）中密切相关。然而，由于缺乏有效的方法来同步大量细胞的细胞周期，因此在分子水平上研究这些相互作用仍然具有挑战性。在这里，我们筛选了细胞周期进程的抑制剂，并确定了诺考达唑是将 hPSCs 同步到 G2/M 期的最有效小分子。诺考达唑处理后，hPSCs 仍保持多能性，保持正常核型，并能成功分化为三个胚层和功能性细胞类型。此外，对同步其细胞周期并向内胚层谱系分化的单细胞进行全基因组转录组分析验证了我们的发现，并表明诺考达唑处理对内胚层分化过程中的基因表达没有影响。因此，我们的同步方法为研究 hPSCs 中的细胞周期机制提供了一种稳健的方法。
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