Department of Life Sciences, University of Bath, Bath, UK.
School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK.
Methods Mol Biol. 2024;2800:11-25. doi: 10.1007/978-1-0716-3834-7_2.
Fibroblasts are the major producers of the extracellular matrix and regulate its organization. Aberrant signaling in diseases such as fibrosis and cancer can impact the deposition of the matrix proteins, which can in turn act as an adhesion scaffold and signaling reservoir promoting disease progression. To study the composition and organization of the extracellular matrix as well as its interactions with (tumor) cells, this protocol describes the generation and analysis of 3D fibroblast-derived matrices and the investigation of (tumor) cells seeded onto the 3D scaffolds by immunofluorescent imaging and cell adhesion, colony formation, migration, and invasion/transmigration assays.
成纤维细胞是细胞外基质的主要产生者，并调节其组织。在纤维化和癌症等疾病中，异常信号会影响基质蛋白的沉积，而这些蛋白反过来又可以作为黏附支架和信号储存库，促进疾病的进展。为了研究细胞外基质的组成和组织，以及其与（肿瘤）细胞的相互作用，本方案描述了 3D 成纤维细胞衍生基质的生成和分析，以及通过免疫荧光成像和细胞黏附、集落形成、迁移和侵袭/转染测定研究接种在 3D 支架上的（肿瘤）细胞。
