Demystifying the effects of a three-dimensional microenvironment in tissue morphogenesis.

Tissue morphogenesis and homeostasis are dependent on a complex dialogue between multiple cell types and chemical and physical cues in the surrounding microenvironment. The emergence of engineered three-dimensional (3D) tissue constructs and the development of tractable methods to recapitulate the native tissue microenvironment ex vivo has led to a deeper understanding of tissue-specific behavior. However, much remains unclear about how the microenvironment and aberrations therein directly affect tissue morphogenesis and behavior. Elucidating the role of the microenvironment in directing tissue-specific behavior will aid in the development of surrogate tissues and tractable approaches to diagnose and treat chronic-debilitating diseases such as cancer and atherosclerosis. Toward this goal, 3D organotypic models have been developed to clarify the mechanisms of epithelial morphogenesis and the subsequent maintenance of tissue homeostasis. Here we describe the application of these 3D culture models to illustrate how the microenvironment plays a critical role in regulating mammary tissue function and signaling, and discuss the rationale for applying precisely defined organotypic culture assays to study epithelial cell behavior. Experimental methods are provided to generate and manipulate 3D organotypic cultures to study the effect of matrix stiffness and matrix dimensionality on epithelial tissue morphology and signaling. We end by discussing technical limitations of currently available systems and by presenting opportunities for improvement.