当前位置: X-MOL 学术Nat. Methods › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Reduced MEK inhibition preserves genomic stability in naive human embryonic stem cells
Nature Methods ( IF 48.0 ) Pub Date : 2018-08-20 , DOI: 10.1038/s41592-018-0104-1
Bruno Di Stefano , Mai Ueda , Shan Sabri , Justin Brumbaugh , Aaron J. Huebner , Anna Sahakyan , Kendell Clement , Katie J. Clowers , Alison R. Erickson , Keiko Shioda , Steven P. Gygi , Hongcang Gu , Toshi Shioda , Alexander Meissner , Yasuhiro Takashima , Kathrin Plath , Konrad Hochedlinger

Human embryonic stem cells (hESCs) can be captured in a primed state in which they resemble the postimplantation epiblast, or in a naive state where they resemble the preimplantation epiblast. Naive-cell-specific culture conditions allow the study of preimplantation development ex vivo but reportedly lead to chromosomal abnormalities, which compromises their utility in research and potential therapeutic applications. Although MEK inhibition is essential for the naive state, here we show that reduced MEK inhibition facilitated the establishment and maintenance of naive hESCs that retained naive-cell-specific features, including global DNA hypomethylation, HERVK expression, and two active X chromosomes. We further show that hESCs cultured under these modified conditions proliferated more rapidly; accrued fewer chromosomal abnormalities; and displayed changes in the phosphorylation levels of MAPK components, regulators of DNA damage/repair, and cell cycle. We thus provide a simple modification to current methods that can enable robust growth and reduced genomic instability in naive hESCs.



中文翻译:

减少的MEK抑制保留了人类胚胎干细胞的基因组稳定性

人类胚胎干细胞(hESCs)可以在类似于植入后成胚细胞的启动状态下捕获,也可以在类似于植入前成胚细胞的幼稚状态下捕获。幼稚细胞特有的培养条件允许进行离体植入前发育的研究,但据报道会导致染色体异常,从而损害了它们在研究和潜在治疗应用中的效用。尽管MEK抑制对于原始状态至关重要,但是在这里我们显示出降低的MEK抑制作用促进了建立和维持保留了原始细胞特有特征的原始hESC的存在,这些特征包括全局DNA低甲基化,HERVK表达和两个活跃的X染色体。我们进一步表明,在这些改良条件下培养的hESCs增殖更为迅速。产生较少的染色体异常;并显示出MAPK成分的磷酸化水平,DNA损伤/修复的调节剂和细胞周期的变化。因此,我们对当前方法进行了简单的修改,可以在幼稚的hESC中实现稳健的生长并降低基因组的不稳定性。

更新日期:2018-08-20
down
wechat
bug