生物
倍性
生殖系
胚胎干细胞
单元格排序
体细胞
细胞生物学
遗传学
孤雌生殖
干细胞
细胞周期
细胞
胚胎
基因
作者
Saori Takahashi,Jiyoung Lee,Takashi Kohda,Ayumi Matsuzawa,Miyuri Kawasumi,Masami Kanai‐Azuma,Tomoko Kaneko‐Ishino,Fumitoshi Ishino
出处
期刊:Development
[The Company of Biologists]
日期:2014-09-25
卷期号:141 (20): 3842-3847
被引量:50
摘要
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.
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