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Biochemical and mechanical extracellular matrix properties dictate mammary epithelial cell motility and assembly

Biochemical and mechanical cues of the extracellular matrix have been shown to play important roles in cell‐matrix and cell‐cell interactions. We have experimentally tested the combined influence of these cues to better understand cell motility, force generation, cell‐cell interaction, and assembly in an in vitro breast cancer model. MCF‐10A non‐tumorigenic mammary epithelial cells were observed on surfaces with varying fibronectin ligand concentration and polyacrylamide gel rigidity. Our data show that cell velocity is biphasic in both matrix rigidity and adhesiveness. The maximum cell migration velocity occurs only at specific combination of substrate stiffness and ligand density. We found cell‐cell interactions reduce migration velocity. However, the traction forces cells exert onto the substrate increase linearly with both cues, with cells in pairs exerting higher maximum tractions observed over single cells. A relationship between force and motility shows a maximum in single cell velocity not observed in cell pairs. Cell‐cell adhesion becomes strongly favored on softer gels with elasticity ≤1250 Pascals (Pa), implying the existence of a compliance threshold that promotes cell‐cell over cell‐matrix adhesion. Finally on gels with stiffness similar to pre‐malignant breast tissue, 400 Pa, cells undergo multicellular assembly and division into 3D spherical aggregates on a 2D surface.