Induction of primordial germ cell-like cells from common marmoset embryonic stem cells by inhibition of WNT and retinoic acid signaling

Reconstitution of germ cell lineage using pluripotent stem cells provides a unique platform to deepen our understanding of the mechanisms underlying germ cell development and to produce functional gametes for reproduction. Here, we report a robust induction of primordial germ cell-like cells (PGCLCs) from common marmoset (Callithrix jacchus) embryonic stem cells. The robust induction was achieved by not only activation of the conserved PGC-inducing signals, WNT and BMP4, but also temporal inhibitions of WNT and retinoic acid signals, which prevent mesodermal and neural differentiation, respectively, during PGCLC differentiation. Many of the gene expression and differentiation properties of common marmoset PGCLCs were similar to those of human PGCLCs, making this culture system a reliable and useful primate model. Finally, we identified PDPN and KIT as surface marker proteins by which PGCLCs can be isolated from embryonic stem cells without genetic manipulation. This study will expand the opportunities for research on germ cell development and production of functional gametes to the common marmoset. RNA-seq analysis of common marmoset embryonic stem cells (cjESC), pre-induced cells, differentiated cells to common marmoset primordial germ cell-like cells (cjPGCLC) or others in multiple conditions, and cjPGCLCs aggregated with mouse gonadal somatic cells

利用多能干细胞（pluripotent stem cells）重建生殖细胞谱系，可为深化我们对生殖细胞发育核心机制的理解，并为生殖领域生产功能性配子提供独特的研究平台。本研究报道了从普通狨猴（Callithrix jacchus）胚胎干细胞中高效诱导原始生殖细胞样细胞（primordial germ cell-like cells, PGCLCs）的方法。该高效诱导体系不仅通过激活保守的PGC诱导信号WNT与BMP4实现，还通过时序性抑制WNT及视黄酸信号通路——前者可阻断中胚层分化，后者可抑制神经分化——在PGCLC分化过程中发挥调控作用。普通狨猴PGCLCs的多数基因表达与分化特性与人类PGCLCs高度相似，使得该培养体系成为可靠且实用的非人灵长类研究模型。最后，本研究鉴定出PDPN与KIT作为表面标志物蛋白，可无需遗传操作即可从胚胎干细胞中分离得到PGCLCs。本研究将为普通狨猴的生殖细胞发育研究及功能性配子生产开辟新的机遇。本数据集包含多种培养条件下的普通狨猴胚胎干细胞（cjESC）、预诱导细胞、向普通狨猴原始生殖细胞样细胞（cjPGCLC）分化的细胞及其他相关样本的RNA测序分析，以及与小鼠性腺体细胞共聚集的cjPGCLCs的相关数据。