Reconstitution of the Mouse Germ Cell Specification Pathway in Culture by Pluripotent Stem Cells
Reconstitution of the Mouse Germ Cell Specification Pathway in Culture by Pluripotent Stem Cells
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DOI：
10.1016/j.cell.2011.06.052
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发表时间：
2011-08-19
期刊：
影响因子：
64.5
通讯作者：
Saitou, Mitinori
中科院分区：
文献类型：
--
作者：
Hayashi, Katsuhiko;Ohta, Hiroshi;Saitou, Mitinori
在体外产生功能正常的配子需要重建生殖细胞发育的多步途径。我们在这里展示了原始生殖细胞样细胞(PGCLCs)在具有强大的精子发生能力的小鼠中的产生。PGCLCs是由胚胎干细胞(ESCs)和经上胚样细胞(EpiLCs)诱导的多能干细胞(IPSCs)产生的，这种细胞状态与原肠前成纤维细胞高度相似，但有别于上胚层干细胞(EPISCs)。从ESCs/IPSCs诱导EpiLC是一个渐进的过程，反映了上皮细胞的发育，而具有PGC命运能力的EpiLC是一个暂时性实体。在从EpiLCs诱导PGCLC的过程中，全球转录图谱、表观遗传学重编程和细胞动力学仔细地从上皮细胞中捕获与PGC规范相关的那些。此外，我们确定整合素-β3和SSEA1是允许从形成肿瘤的未分化细胞中分离出具有生精能力的PGCLCs的标记。我们的发现为体外配子发生的第一步提供了一个范例。
The generation of properly functioning gametes in vitro requires reconstitution of the multistepped pathway of germ cell development. We demonstrate here the generation of primordial germ cell-like cells (PGCLCs) in mice with robust capacity for spermatogenesis. PGCLCs were generated from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) through epiblast-like cells (EpiLCs), a cellular state highly similar to pregastrulating epiblasts but distinct from epiblast stem cells (EpiSCs). Reflecting epiblast development, EpiLC induction from ESCs/iPSCs is a progressive process, and EpiLCs highly competent for the PGC fate are a transient entity. The global transcription profiles, epigenetic reprogramming, and cellular dynamics during PGCLC induction from EpiLCs meticulously capture those associated with PGC specification from the epiblasts. Furthermore, we identify Integrin-beta 3 and SSEA1 as markers that allow the isolation of PGCLCs with spermatogenic capacity from tumorigenic undifferentiated cells. Our findings provide a paradigm for the first step of in vitro gametogenesis.