Method to Synchronize Cell Cycle of Human Pluripotent Stem Cells without Affecting Their Fundamental Characteristics
Method to Synchronize Cell Cycle of Human Pluripotent Stem Cells without Affecting Their Fundamental Characteristics
复制标题
DOI：
10.1016/j.stemcr.2018.11.020
复制
发表时间：
2018-12
影响因子：
5.9
通讯作者：
L. Yiangou;R. Grandy;C. Morell;R. Tomaz;Anna Osnato;Juned Kadiwala;D. Muraro;J. Garcia-Bernardo;Shota Nakanoh;W. G. Bernard;Daniel Ortmann;Davis J. McCarthy;I. Simonic;S. Sinha;L. Vallier
中科院分区：
文献类型：
--
作者：
L. Yiangou;R. Grandy;C. Morell;R. Tomaz;Anna Osnato;Juned Kadiwala;D. Muraro;J. Garcia-Bernardo;Shota Nakanoh;W. G. Bernard;Daniel Ortmann;Davis J. McCarthy;I. Simonic;S. Sinha;L. Vallier
在人类多能干细胞（hPSCs）中，细胞周期进程和细胞命运决定密切相关。然而，在分子水平上对这些相互作用的研究仍然具有挑战性，因为缺乏有效的方法允许大量细胞的细胞周期同步。在这里，我们筛选了细胞周期进程的抑制剂，并确定nocodazole是在G2/M期同步hPSCs最有效的小分子。经诺可达唑处理后，人乳头状细胞保持多能性，保持正常核型，并能成功分化为三种胚层和功能细胞类型。此外，对细胞周期同步并向内胚层谱系分化的单细胞的全基因组转录组学分析证实了我们的发现，并表明诺可达唑对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的细胞周期机制提供了一种可靠的方法。
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.