Chapter 17 - Three-Dimensional Cultures of Normal and Malignant Human Breast Epithelial Cells to Achieve in vivo-like Architecture and Function

Abstract

This chapter describes an assay in which primary or phenotypically normal human breast epithelial cells, cultured in a laminin-rich basement membrane (lrBM), undergo a three-dimensional reorganization to form structures that mimic in vivo acinar structures in culture. Primary breast luminal cells from human reduction mammoplasty as well as human mammary epithelial cell lines have been successfully cultured utilizing the 3D BM assay. The HMT3522 progression series consists of immortal human mammary epithelial cells originally isolated from fibrocystic breast tissue and includes the phenotypically normal S1 cells, as well as their tumorigenic derivative T4-2 cells, which were selected for their ability to grow in the absence of EGF. Successful growth of these cells requires special attention to the media, confluency, trypsinization, and feeding regime in order to maintain healthy cells capable of undergoing differentiation. Single phenotypically normal mammary epithelial cells embedded into lrBM will undergo several rounds of cell division, withdraw from the cell cycle between 6 and 8 days depending on the cell type, organize into polarized structures, and form acini-like structures, including a central lumen.