Tian Chenglei, Liu Linlin, Zeng Ming, Sheng Xiaoyan, Heng Dai, Wang Lingling, Ye Xiaoying, Keefe David L, Liu Lin
State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China.
Department of Cell Biology and Genetics, College of Life Sciences, Nankai University, Tianjin, 300071, China.
Protein Cell. 2021 Dec;12(12):947-964. doi: 10.1007/s13238-021-00865-4. Epub 2021 Nov 30.
Parthenogenetic embryos, created by activation and diploidization of oocytes, arrest at mid-gestation for defective paternal imprints, which impair placental development. Also, viable offspring has not been obtained without genetic manipulation from parthenogenetic embryonic stem cells (pESCs) derived from parthenogenetic embryos, presumably attributable to their aberrant imprinting. We show that an unlimited number of oocytes can be derived from pESCs and produce healthy offspring. Moreover, normal expression of imprinted genes is found in the germ cells and the mice. pESCs exhibited imprinting consistent with exclusively maternal lineage, and higher X-chromosome activation compared to female ESCs derived from the same mouse genetic background. pESCs differentiated into primordial germ cell-like cells (PGCLCs) and formed oocytes following in vivo transplantation into kidney capsule that produced fertile pups and reconstituted ovarian endocrine function. The transcriptome and methylation of imprinted and X-linked genes in pESC-PGCLCs closely resembled those of in vivo produced PGCs, consistent with efficient reprogramming of methylation and genomic imprinting. These results demonstrate that amplification of germ cells through parthenogenesis faithfully maintains maternal imprinting, offering a promising route for deriving functional oocytes and having potential in rebuilding ovarian endocrine function.
通过卵母细胞激活和二倍体化产生的孤雌生殖胚胎,因父源印记缺陷而在妊娠中期停滞,这会损害胎盘发育。此外,未经基因操作,从孤雌生殖胚胎衍生的孤雌生殖胚胎干细胞(pESC)尚未获得存活后代,这可能归因于它们异常的印记。我们表明,可以从pESC中获得无限数量的卵母细胞并产生健康后代。此外,在生殖细胞和小鼠中发现了印记基因的正常表达。pESC表现出与仅母系谱系一致的印记,并且与来自相同小鼠遗传背景的雌性ESC相比,X染色体激活更高。pESC分化为原始生殖细胞样细胞(PGCLC),并在体内移植到肾包膜后形成卵母细胞,这些卵母细胞产生可育幼崽并重建卵巢内分泌功能。pESC-PGCLC中印迹和X连锁基因的转录组和甲基化与体内产生的PGC非常相似,这与甲基化和基因组印记的有效重编程一致。这些结果表明,通过孤雌生殖扩增生殖细胞能忠实地维持母源印记,为获得功能性卵母细胞提供了一条有前景的途径,并具有重建卵巢内分泌功能的潜力。