Abstract：
Human primordial germ cell (PGC) development initiates about 2 weeks after fertilization during embryogenesis. Unique molecular events follow, including epigenetic resetting, to establish functional gametes (egg and sperm). Due to the inaccessibility of human embryos, it is essential to have an amenable experimental platform to investigate the mechanisms and potential dysfunctions of the events. We previously established a PGC-like cell (PGCLC) differentiation method using human pluripotent stem cells (PSCs) via induction of precursor cells followed by stimulation with a cytokine cocktail including BMP. We also revealed that the expression of PGC specifiers, SOX17 and PRDM1, can robustly induce PGCLCs from PSCs without the cytokines. The balance of SOX17 and PRDM1 is critical for germ cell fate since the two factors also regulate endoderm differentiation. Here we describe a detailed procedure for PGCLC differentiation with the balanced induction of SOX17 and PRDM1. The protocol can be used for PGC induction in other mammalian species exhibiting PGCs with SOX17 expression. Together, these studies will advance the understanding of germ cell biology and its applications in reproductive technology and medicine.
摘要：
人原始生殖细胞（PGC）的发育在胚胎发生过程中受精后约2周开始。独特的分子事件随之而来，包括表观遗传重置，建立功能性配子（卵子和精子）。由于人类胚胎难以接近,有一个适合的实验平台来研究事件的机制和潜在的功能障碍是至关重要的。我们先前使用人多能干细胞(PSC)通过前体细胞的诱导,随后用包括BMP的细胞因子混合物刺激,建立了PGC样细胞(PGCLC)分化方法。我们还揭示了PGC说明符的表达，SOX17和PRDM1可以在没有细胞因子的情况下从PSC稳健地诱导PGCLC。SOX17和PRDM1的平衡对于生殖细胞命运至关重要，因为这两种因素也调节内胚层分化。在这里，我们描述了通过平衡诱导SOX17和PRDM1进行PGCLC分化的详细程序。该方案可用于在表现出具有SOX17表达的PGCs的其他哺乳动物物种中诱导PGC。一起,这些研究将促进对生殖细胞生物学及其在生殖技术和医学中的应用的理解。
