Ex utero mouse embryogenesis from pre-gastrulation to late organogenesis
Ex utero mouse embryogenesis from pre-gastrulation to late organogenesis
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DOI：
10.1038/s41586-021-03416-3
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发表时间：
2021-03-17
期刊：
影响因子：
64.8
通讯作者：
Hanna, Jacob H.
中科院分区：
文献类型：
--
作者：
Aguilera-Castrejon, Alejandro;Oldak, Bernardo;Hanna, Jacob H.
哺乳动物的身体计划是在胚胎植入母体子宫后不久建立的，我们对植入后发育过程的理解仍然有限。尽管植入前和植入前后的小鼠胚胎常规地在体外培养，但是用于从卵柱阶段直到高级器官发生的植入后胚胎的稳健培养的方法仍有待建立。在这里，我们提出了一个非常有效的平台，用于植入后小鼠胚胎的子宫外培养，使胚胎从原肠胚形成前（胚胎日（E）5.5），直到后肢形成阶段（E11）的适当发展。晚期原肠胚（E7.5）在三维旋转瓶中生长，而从原肠胚形成前阶段（E5.5或E6.5）的延长培养需要静态和旋转瓶培养平台的组合。组织学、分子和单细胞RNA测序分析证实，子宫外培养的胚胎精确地再现了子宫内发育。该培养系统适合于引入各种胚胎扰动和显微操作，其结果可以在子宫外进行长达六天。建立一个从原肠胚形成前到晚期器官发生的正常小鼠胚胎在子宫外健壮生长的系统是研究胚胎发生的一个有价值的工具，因为它消除了子宫屏障，并允许研究人员机械地询问哺乳动物的植入后形态发生和人工胚胎发生。
The mammalian body plan is established shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. Although pre- and peri-implantation mouse embryos are routinely cultured in vitro', approaches for the robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. Here we present highly effective platforms for the ex utero culture of post-implantation mouse embryos, which enable the appropriate development of embryos from before gastrulation (embryonic day (E) 5.5) until the hindlimb formation stage (E11). Late gastrulating embryos (E7.5) are grown in three-dimensional rotating bottles, whereas extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of static and rotating bottle culture platforms. Histological, molecular and single-cell RNA sequencing analyses confirm that the ex utero cultured embryos recapitulate in utero development precisely. This culture system is amenable to the introduction of a variety of embryonic perturbations and micro-manipulations, the results of which can be followed ex utero for up to six days. The establishment of a system for robustlygrowing normal mouse embryos ex utero from pre-gastrulation to advanced organogenesis represents a valuable tool for investigating embryogenesis, as it eliminates the uterine barrier and allows researchersto mechanistically interrogate post-implantation morphogenesis and artificial embryogenesis in mammals.