Induction of the G2/M transition stabilizes haploid embryonic stem cells
Induction of the G2/M transition stabilizes haploid embryonic stem cells
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DOI：
10.1242/dev.110726
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发表时间：
2014-10-01
期刊：
影响因子：
4.6
通讯作者：
Ishino, Fumitoshi
中科院分区：
文献类型：
--
作者：
Takahashi, Saori;Lee, Jiyoung;Ishino, Fumitoshi
近年来，小鼠孤雌单倍体胚胎干细胞（phESCs）和雄激素单倍体胚胎干细胞（ahESCs）的成功建立，不仅在体外，而且由于这些细胞系的生殖系能力，刺激了遗传学研究。然而，由于单倍体细胞容易自我二倍体化，如果不通过荧光激活细胞分选（FACS）在短时间间隔内对G1期单倍体ESCs进行频繁的分选，则很难长期维持单倍体状态。为了克服这种自发的二倍体转化，我们开发了一种使用小分子Wee1激酶抑制剂的phESC培养条件，通过加速G2/M期转变和防止重新进入额外的G1/S期来调节细胞周期。在此，我们证明，在这种条件下，phESCs在不需要FACS的情况下保持单倍体状态至少4周。这种方法将大大提高这些细胞的遗传筛选的可用性。
The recent successful establishment of mouse parthenogenetic haploid embryonic stem cells (phESCs) and androgenetic haploid ESCs (ahESCs) has stimulated genetic research not only in vitro but also in vivo because of the germline competence of these cell lines. However, it is difficult to maintain the haploid status over time without a frequent sorting of the G1 phase haploid ESCs by fluorescence-activated cell sorting (FACS) at short intervals, because haploid cells tend to readily self-diploidize. To overcome this spontaneous diploid conversion, we developed a phESC culture condition using a small molecular inhibitor of Wee1 kinase to regulate the cell cycle by accelerating the G2/M phase transition and preventing re-entry into extra G1/S phase. Here, we demonstrate that, under this condition, phESCs maintained the haploid status for at least 4 weeks without the need for FACS. This method will greatly enhance the availability of these cells for genetic screening.