Abstract：
Neural tube (NT) defects arise from abnormal neurulation and result in the most common birth defects worldwide. Yet, mechanisms of primate neurulation remain largely unknown due to prohibitions on human embryo research and limitations of available model systems. Here, we establish a three-dimensional (3D) prolonged in vitro culture (pIVC) system supporting cynomolgus monkey embryo development from 7 to 25 days post-fertilization. Through single-cell multi-omics analyses, we demonstrate that pIVC embryos form three germ layers, including primordial germ cells, and establish proper DNA methylation and chromatin accessibility through advanced gastrulation stages. In addition, pIVC embryo immunofluorescence confirms neural crest formation, NT closure, and neural progenitor regionalization. Finally, we demonstrate that the transcriptional profiles and morphogenetics of pIVC embryos resemble key features of similarly staged in vivo cynomolgus and human embryos. This work therefore describes a system to study non-human primate embryogenesis through advanced gastrulation and early neurulation.
摘要：
神经管（NT）缺陷是由神经异常引起的，并导致全球最常见的出生缺陷。然而，由于禁止人类胚胎研究和可用模型系统的限制，灵长类动物神经化的机制在很大程度上仍然未知。这里,我们建立了一个三维（3D）延长体外培养（pIVC）系统，支持食蟹猴胚胎发育7至25天受精后。通过单细胞多组学分析，我们证明pIVC胚胎形成三个胚层，包括原始生殖细胞，并通过高级胃泌素阶段建立适当的DNA甲基化和染色质可及性。此外,pIVC胚胎免疫荧光证实了神经c的形成，NT闭合,和神经祖细胞区域化。最后,我们证明pIVC胚胎的转录谱和形态遗传学类似于体内类似阶段的食蟹猴和人类胚胎的关键特征.因此，这项工作描述了一种通过高级原肠胚形成和早期神经形成研究非人灵长类胚胎发生的系统。
