scRNA-seq analysis of in vitro cultured monkey embryos

Primates embryo implants into the maternal uterus, forms three germ layers and begin early organogenesis, such as neurulation, heart formation, during the first 4 weeks of development. Although, this stage represents a milestone in early embryonic development of primate, our understanding remains limited. Here, we optimized our previously established in vitro culture system, which enable the ex-utero culture of cynomolgus monkey up to 25 days. Morphology histological, and single-cell RNA sequencing analyses confirm that the in vitro cultured embryos recapitulate key events of in vivo development. Using this platform, we delineate monkey early-stage lineage trajectory and genetic programs accompanying the neural, mesoderm and endoderm specification, as well as primordial germ cell–like cell (PGCLC). Thus, this culture system represents a valuable and easily accessible platform to study and manipulate early primate embryogenesis, which cannot easily accomplish in vivo. single cell RNA sequencing of in vitro cultured monkey embryos were performed using 10x Genomics Chromium

灵长类胚胎植入母体子宫后，会形成三胚层并启动早期器官发生过程，例如神经胚形成、心脏形成，该过程发生于发育的前4周。尽管该阶段是灵长类早期胚胎发育的重要里程碑，但目前学界对其认知仍十分有限。本研究优化了此前建立的体外培养体系，该体系可支持食蟹猴（cynomolgus monkey）胚胎的子宫外培养至第25天。形态学、组织学及单细胞RNA测序（single-cell RNA sequencing）分析证实，体外培养的胚胎能够重现体内发育的关键事件。借助该平台，我们解析了食蟹猴早期谱系发育轨迹，以及伴随神经胚、中胚层、内胚层特化与原始生殖细胞样细胞（primordial germ cell-like cell, PGCLC）形成的基因调控程序。综上，该培养体系为研究与操控早期灵长类胚胎发生提供了极具价值且易于获取的平台——这一研究在体内实验中难以实现。本研究使用10x Genomics Chromium平台完成了体外培养食蟹猴胚胎的单细胞RNA测序。