mRNA-based generation of marmoset PGCLCs capable of differentiation into gonocyte-like cells
mRNA-based generation of marmoset PGCLCs capable of differentiation into gonocyte-like cells
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DOI：
10.1101/2022.09.20.508677
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发表时间：
2022-09
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通讯作者：
Musashi Kubiura-Ichimaru;Christopher A. Penfold;Kazuaki Kojima;Constance Dollet;Haruka Yabukami;Katsunori Semi;Yasuhiro Takashima;Thorsten E. Boroviak;H. Kawaji;K. Woltjen;Aki Minoda;E. Sasaki;Toshiaki Watanabe
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作者：
Musashi Kubiura-Ichimaru;Christopher A. Penfold;Kazuaki Kojima;Constance Dollet;Haruka Yabukami;Katsunori Semi;Yasuhiro Takashima;Thorsten E. Boroviak;H. Kawaji;K. Woltjen;Aki Minoda;E. Sasaki;Toshiaki Watanabe
由于样本收集的限制和发育的长时间，灵长类生殖细胞的发育在很大程度上仍未被探索。在小鼠中，多能干细胞（PSCs）衍生的原始生殖细胞样细胞（pgclc）可以通过体外培养或体内移植发育成功能配子。这种pgclc介导的灵长类动物成熟配子的诱导对理解人类生殖细胞的发育非常有用。由于狨猴比其他物种更早产生有功能的精子，概括整个雄性生殖细胞的发育过程在技术上更可行。在这里，我们通过绒猴的pgclc诱导iPSCs向淋细胞样细胞分化。首先，我们开发了一种基于mRNA转染的方法来高效地生成PGCLC。随后，为了促进PGCLC分化，在小鼠肾包膜中生成异种重构睾丸（xrtestes）。pgclc表现出进行性DNA去甲基化和发育标记基因的逐步表达。本研究为绒猴生殖细胞发育的研究提供了一个有效的平台。本文综述了小鼠肾脏中绒猴PGCLC分化为性腺细胞样细胞的发育调控表达和去甲基化过程。[导语]Watanabe等人利用基于mRNA转染的方法高效诱导绒猴原始生殖细胞样细胞（PGCLC）。pgclc在小鼠肾囊构建的异种重构睾丸中进一步发展为淋细胞样细胞。该系统忠实地再现了体内发育过程（例如，发育调节基因的阶段特异性表达和DNA去甲基化）。
Primate germ cell development remains largely unexplored due to limitations in sample collection and the long duration of development. In mice, primordial germ cell-like cells (PGCLCs) derived from pluripotent stem cells (PSCs) can develop into functional gametes by in vitro culture or in vivo transplantation. Such PGCLC-mediated induction of mature gametes in primates is highly useful for understanding human germ cell development. Since marmosets generate functional sperm earlier than other species, recapitulating the whole male germ cell development process is technically more feasible. Here, we induced the differentiation of iPSCs into gonocyte-like cells via PGCLCs in marmosets. First, we developed an mRNA transfection-based method to efficiently generate PGCLC. Subsequently, to promote PGCLC differentiation, xenoreconstituted testes (xrtestes) were generated in the mouse kidney capsule. PGCLCs show progressive DNA demethylation and stepwise expression of developmental marker genes. This study provides an efficient platform for the study of marmoset germ cell development. Highlights An mRNA transfection-based PGCLC induction method is developed Marmoset PGCLCs are efficiently induced from iPSCs Marmoset PGCLCs differentiate into gonocyte-like cells in mouse kidneys Developmentally regulated expression and demethylation are recapitulated eTOC blurb Watanabe et al. efficiently induced marmoset primordial germ cell-like cells (PGCLCs) using an mRNA transfection-based approach. PGCLCs further develop into gonocyte-like cells in the xenoreconstituted testes constructed under mouse kidney capsules. This system faithfully reproduced in vivo developmental processes (e.g., stage-specific expression of developmentally regulated genes and DNA demethylation).