Investigating Cell-ECM Interactions and ECM Synthesis in Three-Dimensional Hydrogels

Abstract

Model systems composed of cells cultured within three-dimensional (3D) gels are widely used to examine mechanisms of morphogenesis and repair and are applied as vehicles for delivering cells for therapeutic applications in vivo. Cells secrete extracellular matrix (ECM), the composition of which depends on the type of cell and cues present; the ECM is assembled and remodeled, forming a dynamic cellular niche. While it is generally recognized that the presence of ECM and cellular interactions with the ECM, for example via integrin binding, are closely tied to cell and tissue behavior, few investigations have centered on quantifying these processes in 3D gel microenvironments. Herein, we provide methods to characterize the location and patterns of ECM and cell-ECM interactions using immunofluorescence and collect cell lysate for analysis via immunoblotting or ELISA.