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从多能干细胞中诱导人类生殖细胞命运的稳健体外方法。

Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells.

机构信息

Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; JST, ERATO, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.

Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; JST, ERATO, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.

出版信息

Cell Stem Cell. 2015 Aug 6;17(2):178-94. doi: 10.1016/j.stem.2015.06.014. Epub 2015 Jul 16.

Abstract

Mechanisms underlying human germ cell development are unclear, partly due to difficulties in studying human embryos and lack of suitable experimental systems. Here, we show that human induced pluripotent stem cells (hiPSCs) differentiate into incipient mesoderm-like cells (iMeLCs), which robustly generate human primordial germ cell-like cells (hPGCLCs) that can be purified using the surface markers EpCAM and INTEGRINα6. The transcriptomes of hPGCLCs and primordial germ cells (PGCs) isolated from non-human primates are similar, and although specification of hPGCLCs and mouse PGCs rely on similar signaling pathways, hPGCLC specification transcriptionally activates germline fate without transiently inducing eminent somatic programs. This includes genes important for naive pluripotency and repression of key epigenetic modifiers, concomitant with epigenetic reprogramming. Accordingly, BLIMP1, which represses somatic programs in mice, activates and stabilizes a germline transcriptional circuit and represses a default neuronal differentiation program. Together, these findings provide a foundation for understanding and reconstituting human germ cell development in vitro.

摘要

人类生殖细胞发育的机制尚不清楚,部分原因是难以研究人类胚胎和缺乏合适的实验系统。在这里,我们表明,人类诱导多能干细胞(hiPSCs)分化为初始中胚层样细胞(iMeLCs),这些细胞可以强烈地产生类似于人类原始生殖细胞样细胞(hPGCLCs),可以使用表面标记物 EpCAM 和 INTEGRINα6 进行纯化。从非人类灵长类动物中分离的 hPGCLCs 和原始生殖细胞(PGCs)的转录组相似,尽管 hPGCLCs 和小鼠 PGCs 的特化依赖于类似的信号通路,但 hPGCLC 的特化转录激活生殖系命运,而不会短暂诱导突出的体细胞程序。这包括对原始多能性和关键表观遗传修饰物抑制的重要基因,伴随着表观遗传重编程。相应地,BLIMP1 在小鼠中抑制体细胞程序,激活并稳定生殖系转录电路,并抑制默认的神经元分化程序。总之,这些发现为理解和体外重建人类生殖细胞发育提供了基础。

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