SummaryThe generation of germ cells from embryonic stem cells in vitro has current historical significance. Western blot, qPCR, immunofluorescence and flow cytometry assays were used to investigate the differences in expression levels of totipotency and specific markers for Wnt regulation and the related signalling pathways during primordial germ cell-like cell (PGCLC) induction and differentiation. During PGCLC induction, activation of WNT3a increased the expression of NANOG, SOX2 and OCT4, but Mvh, DAZL, Blimp1, TFAP2C, Gata4, SOX17, EOMES, Brachyury and PRDM1 expression levels were significantly reduced. Inhibition of the WNT signal demonstrated the opposite effect. Similarly, inhibitors of BMP and the Nodal/Activin signal were used to determine the effect of signal pathways on differentiation. CER1 affected the Wnt signal and differentiation, but the inhibitor SB only regulated differentiation. BMP–WNT–NODAL were mainly responsible for regulating differentiation. Our results provide a reliable theoretical basis and feasibility for further clinical medical research.

中文翻译：

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BMP-Wnt-Nodal 信号对干细胞分化的影响

摘要从胚胎干细胞产生生殖细胞体外具有当今的历史意义。使用蛋白质印迹、qPCR、免疫荧光和流式细胞术分析来研究原始生殖细胞样细胞 (PGCLC) 诱导和分化过程中全能性和 Wnt 调控的特异性标志物以及相关信号通路的表达水平的差异。在 PGCLC 诱导期间，WNT3a 的激活增加了 NANOG、SOX2 和 OCT4 的表达，但 Mvh、DAZL、Blimp1、TFAP2C、Gata4、SOX17,EOMES,布拉丘里和PRDM1表达水平显着降低。WNT 信号的抑制表现出相反的效果。类似地，使用 BMP 抑制剂和 Nodal/Activin 信号来确定信号通路对分化的影响。CER1 影响 Wnt 信号和分化，但抑制剂 SB 仅调节分化。BMP-WNT-NODAL 主要负责调节分化。我们的研究结果为进一步的临床医学研究提供了可靠的理论依据和可行性。