IRIBHM, Université Libre de Bruxelles, Brussels, Belgium.
WELBIO, Wavre, Belgium.
Methods Mol Biol. 2021;2179:135-144. doi: 10.1007/978-1-0716-0779-4_12.
Epithelial-mesenchymal transition (EMT) is often studied in pathological contexts, such as cancer or fibrosis. This chapter focuses on physiological EMT that allows the separation of germ layers during mouse embryo gastrulation. In order to record individual cells behavior with high spatial and temporal resolution live imaging as they undergo EMT, it is very helpful to label the cells of interest in a mosaic fashion so as to facilitate cell segmentation and quantitative image analysis. This protocol describes the isolation, culture, and live imaging of E6.5-E7.5 mouse embryos mosaically labeled in the epiblast, the epithelium from which mesoderm and endoderm layers arise through EMT at gastrulation.
上皮-间充质转化（EMT）通常在病理环境中进行研究，如癌症或纤维化。本章重点介绍了在胚胎发生期间允许胚层分离的生理 EMT。为了在经历 EMT 时以高时空分辨率对单个细胞的行为进行实时成像，以马赛克方式标记感兴趣的细胞非常有帮助，这有助于细胞分割和定量图像分析。本协议描述了通过 EMT 在胚盘中产生中胚层和内胚层的上皮细胞的 E6.5-E7.5 鼠标胚胎的分离、培养和马赛克标记的活体成像。
