Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells
Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells
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DOI：
10.1016/j.stem.2015.06.014
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发表时间：
2015-08-06
期刊：
影响因子：
23.9
通讯作者：
Saitou, Mitinori
中科院分区：
文献类型：
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作者：
Sasaki, Kotaro;Yokobayashi, Shihori;Saitou, Mitinori
人类生殖细胞发育的机制尚不清楚，部分原因是研究人类胚胎的困难和缺乏合适的实验系统。在这里，我们展示了人类诱导多能干细胞 (hiPSC) 分化为早期中胚层样细胞 (iMeLC)，后者能够有力地生成人类原始生殖细胞样细胞 (hPGCLC)，并且可以使用表面标记 EpCAM 和 INTEGRIN α 6 进行纯化。从非人类灵长类动物中分离的 hPGCLC 和原始生殖细胞 (PGC) 的转录组 相似，尽管 hPGCLC 和小鼠 PGC 的特异化依赖于相似的信号传导途径，但 hPGCLC 特异化会转录激活种系命运，而不会短暂诱导显着的体细胞程序。这包括对幼稚多能性和关键表观遗传修饰因子的抑制很重要的基因，伴随着表观遗传重编程。因此，抑制小鼠体细胞程序的 BLIMP1 激活并稳定种系转录回路，并抑制默认的神经元分化程序。总之，这些发现为理解和重建人类生殖细胞体外发育奠定了基础。
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 INTEGRIN alpha 6. 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.