Mammary gland is composed of branched epithelial structure embedded within a complex stroma formed by several stromal cell types, including fibroblasts, and extracellular matrix (ECM). Development of mammary gland is tightly regulated by bidirectional epithelial-stromal interactions that include paracrine signaling, ECM remodeling and mechanosensing. Importantly, these interactions play crucial role in mammary gland homeostasis and when deregulated they contribute to tumorigenesis. Therefore, understanding the mechanisms underlying epithelial-stromal interactions is critical for elucidating regulation of normal mammary gland development and homeostasis and revealing novel strategies for breast cancer therapy. To this end, several three-dimensional (3D) cell culture models have been developed to study these interactions in vitro. In this chapter, a novel 3D organoid-fibrosphere coculture model of mammary gland is described with the capacity for studying not only the qualitative and quantitative aspects of interactions between mammary fibroblasts and epithelial organoids but also their radius and directionality.
Keywords: 3D culture; Branching morphogenesis; Collagen; Extracellular matrix; Fibroblasts; Mammary gland; Matrigel; Organoids; Paracrine signaling; Spheroid.