体外培养小鼠囊胚至囊胚腔阶段通过壁滋养层细胞切除。

In Vitro Culture of Mouse Blastocysts to the Egg Cylinder Stage via Mural Trophectoderm Excision.

机构信息

Embryonic Self-Organization Research Group, Max Planck Institute for Molecular Biomedicine, Münster, Germany.

出版信息

Methods Mol Biol. 2021;2214:31-40. doi: 10.1007/978-1-0716-0958-3_3.

Abstract

The developmental transition from the blastocyst to the egg cylinder stage is associated with stark changes in the overall shape of the embryo, as well as with reorganization of the transcriptional network and epigenetic landscape in the pluripotent and the supportive extraembryonic lineages. To directly analyze this pre- to postimplantation switch, culture conditions are needed that can support mouse embryogenesis beyond the blastocyst stage without maternal input. Here we provide a step-by-step protocol describing an experimental pipeline for isolating late blastocysts, excising (manually or via laser assistance) the mural trophectoderm, and, finally, culturing the embryo to the egg cylinder stage.

摘要

从囊胚到胚泡阶段的发育转变与胚胎整体形状的明显变化有关，以及多能性和支持性胚外谱系中转录网络和表观遗传景观的重新组织。为了直接分析这种植入前到植入后的转变，需要培养条件来支持小鼠胚胎发生，使其超越囊胚阶段而无需母体输入。在这里，我们提供了一个逐步的方案，描述了一个从晚期囊胚中分离、切除（手动或通过激光辅助）壁滋养层，最后培养到胚泡阶段的实验流程。