Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
Laboratory of Genetics and Developmental Biology, Institut Curie, INSERM U934, CNRS UMR3215, Paris, France.
Methods Mol Biol. 2024;2764:107-129. doi: 10.1007/978-1-0716-3674-9_8.
Fibroblasts are an integral cell type of mammary gland stroma, which plays crucial roles in development, homeostasis, and tumorigenesis of mammary epithelium. Fibroblasts produce and remodel extracellular matrix proteins and secrete a plethora of paracrine signals, which instruct both epithelial and other stromal cells of the mammary gland through mechanisms, which have not been fully understood. To enable deciphering of the intricate fibroblast-epithelial interactions, we developed several 3D co-culture methods. In this chapter, we describe methods for establishment of various types of embedded 3D co-cultures of mammary fibroblasts with mammary epithelial organoids, mammary tumor organoids, or breast cancer spheroids to investigate the role of fibroblasts in mammary epithelial development, morphogenesis, and tumorigenesis. The co-culture types include dispersed, aggregated, and transwell cultures.
成纤维细胞是乳腺基质的重要细胞类型，在乳腺上皮的发育、稳态和肿瘤发生中发挥着关键作用。成纤维细胞产生和重塑细胞外基质蛋白，并分泌大量旁分泌信号，通过尚未完全理解的机制来指导乳腺的上皮细胞和其他基质细胞。为了能够解析复杂的成纤维细胞-上皮细胞相互作用，我们开发了几种 3D 共培养方法。在本章中，我们描述了建立各种类型的嵌入 3D 共培养物的方法，这些共培养物将乳腺成纤维细胞与乳腺上皮类器官、乳腺肿瘤类器官或乳腺癌球体混合，以研究成纤维细胞在乳腺上皮发育、形态发生和肿瘤发生中的作用。共培养类型包括分散、聚集和 Transwell 培养。
