Clonal variation of human induced pluripotent stem cells for induction into the germ cell fate†
Clonal variation of human induced pluripotent stem cells for induction into the germ cell fate†
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用于诱导生殖细胞命运的人类诱导多能干细胞的克隆变异†
DOI：
10.1093/biolre/iox038
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发表时间：
2017
影响因子：
3.6
通讯作者：
Saitou Mitinori
中科院分区：
文献类型：
--
作者：
Yokobayashi Shihori;Okita Keisuke;Nakagawa Masato;Nakamura Tomonori;Yabuta Yukihiro;Yamamoto Takuya;Saitou Mitinori
由于难以获得合适的实验材料，人类生殖细胞发育的机制在很大程度上仍然未知。因此，建立一个体外系统，以重建人类生殖细胞的发展将提供一个关键的机会，了解其机制在分子水平上。先前已经显示，人诱导多能干细胞（hiPSC）首先被诱导成初始中胚层样细胞（iMeLC），iMeLC继而被诱导成具有与早期迁移性PGC相似的基因表达特性的原始生殖细胞样细胞（PGCLC）。在这里，我们报告了PGCLC诱导的效率在hiPSC克隆之间变化，并且有趣的是，PGCLC诱导效率的克隆变化反映在iMeLC的基因表达状态中。值得注意的是，EOMES、MIXL1或Tin在iMeLCs中的表达水平与随后PGCLC生成的效率正相关，而CDH1、SOX 3或FGF 2的表达水平则呈负相关。这些结果表明，在iMeLC诱导期间发生的这些基因的表达变化是指示PGCLC成功诱导的关键标志物，此外，hiPSC克隆具有影响其对iMeLC诱导的响应性的不同性质。因此，我们的研究提供了重要的见解hPGC的规范机制，以及一个更好的策略，在体外诱导人生殖细胞的命运从PSC的发展。
The mechanisms for human germ cell development have remained largely unknown, due to the difficulty in obtaining suitable experimental materials. The establishment of an in vitro system to reconstitute human germ cell development will thus provide a critical opportunity to understand its mechanisms at a molecular level. It has previously been shown that human induced pluripotent stem cells (hiPSCs) are first induced into incipient mesoderm-like cells (iMeLCs), which are in turn induced into primordial germ-cell like cells (PGCLCs) with gene expression properties similar to early migratory PGCs. Here, we report that the efficiency of PGCLC induction varies among hiPSC clones, and, interestingly, the clonal variations in PGCLC induction efficiency are reflected in the gene expression states of the iMeLCs. Remarkably, the expression levels ofEOMES, MIXL1, orTin the iMeLCs are positively correlated with the efficiency of subsequent PGCLC generation, while the expressions ofCDH1, SOX3, orFGF2are negatively correlated. These results indicate that the expression changes of these genes occurring during iMeLC induction are key markers indicative of successful induction of PGCLCs, and furthermore, that hiPSC clones have different properties that influence their responsivity to the iMeLC induction. Our study thus provides important insights into the mechanism of hPGC specification as well as the development of a better strategy for inducing human germ cell fate from PSCs in vitro.