Setup of an In Vitro Three-Dimensional Stromalized Prostate Cancer Model Using Gelatin Microparticles.

Developing three-dimensional (3D) tumor models that accurately mimic the tumor microenvironment (TME) and its heterogeneity remains a significant challenge in preclinical research. Advancing these models holds the potential to improve the study of cancer pathologies in vitro, while reducing dependence on animal models. To tackle this challenge, in this work, we report on the development of an in vitro 3D stromalized prostate cancer model using gelatin porous microparticles as microscaffolds for cell attachment and growth. Gelatin porous microparticles were prepared by a double emulsion method and cross-linked with a biocompatible cross-linking agent, that is, glyceraldehyde, to prevent dissolution under physiological conditions. Then, we developed a stromalized 3D gelatin-based microscaffold biomimicking the interplay between human prostate cancer (PCa) and stromal cells by coculturing 22Rv1 cells and fibroblasts with gelatin porous microparticles. Overall, our results demonstrate the feasibility of gelatin microscaffolds in reproducing a 3D stromalized model of PCa progression (e.g., metabolic reprogramming), resulting from the tumor-stroma interaction. Thus, these systems represent a valuable platform and an effective tool for the study of cancer progression, such as TME biomimetics, while simultaneously offering a valid alternative to minimize the reliance on animal studies in preclinical research.