In vitro long-term culture of cynomolgus monkey embryos to early neurulation stage (maca_RNA_rawdata)

Neurulation is the process of forming the neural tube. The complex characteristics of primate neurulation remain mysterious due to the limited accessibility of primate embryos at this stage. Here, we established a 3D prolonged in vitro culture (pIVC) system to support the cynomolgus monkey blastocyst developing to E25 to early neurulation. The pIVC E25 embryos highly recapitulated the critical events of their in vivo counterparts during early neurulation, which include the specialization of the neural plate, closure of the neural tube, regionalization of neural tube cells, specialization and migration of neural crest cells and differentiation of motor neurons and sensory placodes. Single-cell multi-omics analysis of the pIVC embryos further verified the specification of early neural cells, suggesting the pIVC embryos developing to a more advanced stage. Besides, the DNA methylation and chromosome accessibility of three germ layers were in-deep analyzed. This study will help to explore the characteristics and underlying mechanisms of primate neurogenesis, which will be of fundamental and clinical importance.

神经胚形成（Neurulation）是神经管形成的过程。由于该阶段灵长类胚胎的可及性有限，灵长类神经胚形成的复杂特征仍未被完全阐明。本研究建立了一种三维长期体外培养（3D prolonged in vitro culture, pIVC）系统，可支持食蟹猴囊胚发育至E25（早期神经胚形成）阶段。该pIVC培养的E25胚胎高度重现了体内对应阶段的关键事件，包括神经板特化、神经管闭合、神经管细胞区域化、神经嵴细胞的特化与迁移，以及运动神经元与感觉基板的分化。对pIVC胚胎的单细胞多组学分析进一步验证了早期神经细胞的特化进程，表明该培养体系可支持胚胎发育至更成熟的阶段。此外，本研究还深入分析了三胚层的DNA甲基化与染色质可及性。本研究有助于揭示灵长类神经发生的特征与潜在机制，具有重要的基础研究与临床转化价值。