生物
表观遗传学
DNA甲基化
胚胎干细胞
发育生物学
干细胞
甲基化
基因组印记
差异甲基化区
倍性
遗传学
细胞生物学
胚胎
人口
作者
Hongling Zhang,Yuanyuan Li,Yu Ma,Chongping Lai,Qian Yu,Guangyong Shi,Jinsong Li
标识
DOI:10.1007/s13238-021-00890-3
摘要
Abstract The use of two inhibitors of Mek1/2 and Gsk3β (2i) promotes the generation of mouse diploid and haploid embryonic stem cells (ESCs) from the inner cell mass of biparental and uniparental blastocysts, respectively. However, a system enabling long-term maintenance of imprints in ESCs has proven challenging. Here, we report that the use of a two-step a2i (alternative two inhibitors of Src and Gsk3β, TSa2i) derivation/culture protocol results in the establishment of androgenetic haploid ESCs (AG-haESCs) with stable DNA methylation at paternal DMRs (differentially DNA methylated regions) up to passage 60 that can efficiently support generating mice upon oocyte injection. We also show coexistence of H3K9me3 marks and ZFP57 bindings with intact DMR methylations. Furthermore, we demonstrate that TSa2i-treated AG-haESCs are a heterogeneous cell population regarding paternal DMR methylation. Strikingly, AG-haESCs with late passages display increased paternal-DMR methylations and improved developmental potential compared to early-passage cells, in part through the enhanced proliferation of H19 -DMR hypermethylated cells. Together, we establish AG-haESCs that can long-term maintain paternal imprints.
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