Development and characterization of a recombinant silk network for 3D culture of immortalized and fresh tumor‐derived breast cancer cells

Traditional cancer models rely on 2D cell cultures or 3D spheroids, which fail to recapitulate cell‐extracellular matrix (ECM) interactions, a key element of tumor development. Existing hydrogel‐based 3D alternatives lack mechanical support for cell growth and often suffer from low reproducibility. Here we report a novel strategy to make 3D models of breast cancer using a tissue‐like, well‐defined network environment based on recombinant spider silk, functionalized with a cell adhesion motif from fibronectin (FN‐silk). With this approach, the canonical cancer cells SK‐BR‐3, MCF‐7, and MDA‐MB‐231, maintain their characteristic expression of markers (i.e., ERα, HER2, and PGR) while developing distinct morphology. Transcriptomic analyses demonstrate how culture in the FN‐silk networks modulates the biological processes of cell adhesion and migration while affecting physiological events involved in malignancy, such as inflammation, remodeling of the ECM, and resistance to anticancer drugs. Finally, we show that integration in FN‐silk networks promotes the viability of cells obtained from the superficial scraping of patients' breast tumors.