Differentiation of Fibroblasts to Adipocytes in 3D for a Co-culture with Mammary Organoids and Immunohistological Analysis

Mammary epithelial ducts, the main functional compartment of the mammary gland, are embedded in an adipocyte-rich stroma, which is essential for proper mammary gland development, function, and tissue homeostasis. Moreover, the adipocyte compartment has an important role in cancer progression. To better understand cell-to-cell interactions and the role of the adipocytes in the mammary gland, development of proper in vitro models which realistically mimic in vivo conditions has been essential. In this chapter, we describe a simple and effective method for generating mammary gland adipocytes from mammary fibroblasts and their subsequent co-culture with mammary epithelial organoids to further investigate the role of adipocytes in epithelial development and morphogenesis.