Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge CB2 0AW, UK.
Guangzhou Institutes of Biomedicine and Health (GIBH), Chinese Academy of Sciences, Guangzhou 510530, China; Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge CB2 0AW, UK.
Stem Cell Reports. 2019 Dec 10;13(6):1083-1098. doi: 10.1016/j.stemcr.2019.10.009. Epub 2019 Nov 7.
In contrast to conventional human pluripotent stem cells (hPSCs) that are related to post-implantation embryo stages, naive hPSCs exhibit features of pre-implantation epiblast. Naive hPSCs are established by resetting conventional hPSCs, or are derived from dissociated embryo inner cell masses. Here we investigate conditions for transgene-free reprogramming of human somatic cells to naive pluripotency. We find that Wnt inhibition promotes RNA-mediated induction of naive pluripotency. We demonstrate application to independent human fibroblast cultures and endothelial progenitor cells. We show that induced naive hPSCs can be clonally expanded with a diploid karyotype and undergo somatic lineage differentiation following formative transition. Induced naive hPSC lines exhibit distinctive surface marker, transcriptome, and methylome properties of naive epiblast identity. This system for efficient, facile, and reliable induction of transgene-free naive hPSCs offers a robust platform, both for delineation of human reprogramming trajectories and for evaluating the attributes of isogenic naive versus conventional hPSCs.
与涉及着床后胚胎阶段的传统人类多能干细胞 (hPSC) 不同，原始 hPSC 表现出着床前上胚层的特征。原始 hPSC 是通过重置传统 hPSC 或从分离的胚胎内细胞团衍生而来的。在这里，我们研究了无转基因的人类体细胞重编程为原始多能性的条件。我们发现 Wnt 抑制促进 RNA 介导的原始多能性诱导。我们证明了在独立的人成纤维细胞培养物和内皮祖细胞中的应用。我们表明，诱导的原始 hPSC 可以在二倍体核型下进行克隆扩增，并在形成性转变后进行体细胞谱系分化。诱导的原始 hPSC 系表现出原始上胚层特征的独特表面标志物、转录组和甲基组特性。这种高效、简便、可靠的无转基因原始 hPSC 诱导系统为描绘人类重编程轨迹和评估同基因原始与传统 hPSC 的特性提供了一个强大的平台。
