Analysis of MCF10A mammary epithelial cell acinar morphogenesis within well-defined 3-dimensinal systems

Epithelial tissue morphogenesis proceeds within the context of a three dimensional (3D) extracellular matrix (ECM). Accordingly, to clarify the molecular basis of tissue-specific differentiation and disease, a variety of 3D systems exploiting natural ECMs have been developed, such as reconstituted basement membrane (rBM) and purified collagen hydrogels. These natural hydrogels recapitulate epithelial tissue architecture and behaviors in vitro with reasonable fidelity. Nevertheless, natural matrices suffer from considerable preparation variability and remain poorly defined biochemically and biophysically. To understand epithelial cell biology requires defined biomaterials in which biochemical, topological and biophysical properties can be systematically varied. Towards this goal we use the nonmalignant MCF10A mammary epithelial cell (MEC) line and are conducting a systematic analysis of acinar morphogenesis using the natural hydrogels collagen type I and rBM and three synthetic matrices: rBM-conjugated polyacrylamide gels, self assembling peptide gels (PuraMatrix) with and without rBM and hyaluronic acid gels with and without rBM. Rigorous, morphometric and quantatitive analysis as well as immunohistochemistry, and electron microscopy are in progress.