Gillich Astrid, Bao Siqin, Surani M Azim
Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2013;1074:15-29. doi: 10.1007/978-1-62703-628-3_2.
Mouse postimplantation epiblast cultured in activin and basic fibroblast growth factor gives rise to continuously growing epiblast stem cells (EpiSCs) that share key properties with postimplantation epiblast, such as DNA methylation and an inactive X-chromosome. EpiSCs also show a distinct gene expression profile compared to embryonic stem cells (ESCs) derived from preimplantation blastocysts, and do not contribute efficiently to chimeras. EpiSCs can, however, revert to pluripotent ESC-like cells upon exposure to leukemia inhibitory factor-Stat3 signalling on feeder cells. Here we describe a protocol for the establishment of EpiSCs and their reversion to ESCs.
在激活素和碱性成纤维细胞生长因子中培养的小鼠植入后上胚层可产生持续生长的上胚层干细胞(EpiSCs),这些细胞与植入后上胚层具有共同的关键特性,如DNA甲基化和失活的X染色体。与源自植入前囊胚的胚胎干细胞(ESCs)相比,EpiSCs也表现出独特的基因表达谱,并且对嵌合体的贡献效率不高。然而,EpiSCs在饲养细胞上暴露于白血病抑制因子-Stat3信号时可恢复为多能性的ESC样细胞。在这里,我们描述了一种建立EpiSCs及其向ESCs逆转的方案。