Centre de Biologie du Développement (CBD), Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, Toulouse Cedex 09, France.
Methods Mol Biol. 2021;2179:257-274. doi: 10.1007/978-1-0716-0779-4_20.
The epithelial-mesenchymal transition (EMT) converts coherent epithelial structures into single cells. EMT is a dynamic cellular process that is not systematically completed (not all EMTs lead to single cells) and reversible (cells can re-epithelialize). EMT is orchestrated at multiple levels from transcription, to posttranslational modifications, to protein turnover. It involves remodeling of polarity and adhesion and enhances migratory capabilities. During physiological events such as embryogenesis or wound healing EMT is used to initiate cell migration, but EMT can also occur in pathological settings. In particular, EMT has been linked to fibrosis and cancer. Neural crest (NC) cells, an embryonic stem cell population whose behavior recapitulates the main steps of carcinoma progression, are a great model to study EMT. In this chapter, we provide a fully detailed protocol to extract NC cells from Xenopus embryos and culture them to study the dynamics of cell-cell adhesion, cell motility, and dispersion.
上皮-间充质转化 (EMT) 将连贯的上皮结构转化为单个细胞。EMT 是一个动态的细胞过程，不会系统地完成（并非所有 EMT 都会导致单个细胞），并且是可逆的（细胞可以重新上皮化）。EMT 在多个层面上进行协调，包括转录、翻译后修饰和蛋白质周转。它涉及到极性和黏附的重塑，并增强迁移能力。在胚胎发生或伤口愈合等生理事件中，EMT 用于启动细胞迁移，但 EMT 也可能发生在病理环境中。特别是，EMT 与纤维化和癌症有关。神经嵴 (NC) 细胞是一种胚胎干细胞群体，其行为重现了癌进展的主要步骤，是研究 EMT 的理想模型。在本章中，我们提供了一个完整的详细方案，从 Xenopus 胚胎中提取 NC 细胞并培养它们，以研究细胞-细胞黏附、细胞迁移和分散的动力学。
