Epigenetic integrity of paternal imprints enhances the developmental potential of androgenetic haploid embryonic stem cells

• 作者: Hongling Zhang Yuanyuan Li Yongjian Ma Chongping Lai Qian Yu Guangyong Shi Jinsong Li
• 来源期刊: 蛋白质与细胞
• 年，卷（期）: 2022,（2）
• 所属期刊栏目: RESEARCH ARTICLES
• 页码范围: 102-119
• 页数: 18页
• 语种: 英文

摘要

The use of two inhibitors of Mek1/2 and Gsk3β (2i) pro-motes 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 poten-tial compared to early-passage cells,in part through the enhanced proliferation of H1g-DMR hypermethylated cells.Together,we establish AG-haESCs that can long-term maintain paternal imprints.