Mimicry of Tumor‐Promoting Factors in 3D In Vitro Breast Cancer Models: AI‐Driven Opportunities for Model Design and Biological Data Analysis

ABSTRACT With the growing need to accurately recapitulate tumor pathophysiology in vitro, 3D culture systems have emerged as superior to standard 2D models. 3D cell culture systems overcome key limitations of 2D cultures, including insufficient cell‐to‐cell and cell‐to‐matrix interactions, lack of cellular heterogeneity, and absence of structural complexity. Physiologically relevant modeling of cancer can only be achieved by adequately mimicking critical tumor promoting factors such as hypoxia, cancer stemness, metastatic potential, and drug resistance—features that are effectively captured by in vitro 3D tumor models developed using scaffold‐free or scaffold‐based platforms. Scaffold‐based systems are particularly valuable, as they provide extracellular matrix (ECM)‐mimicking physical, chemical, and spatiotemporal cues, thereby supporting the formation of robust and pertinent 3D in vitro cancer models. With the fact that breast cancer (BC) is widely growing as a life threatening cancer amongst women, advancing 3D in vitro breast cancer models is crucial for developing biologically relevant and reliable platforms to evaluate drug candidates and treatment regimens. Such platforms offer significant potential for the effective translation of anticancer therapies and personalized medicine. This perspective discusses scaffold‐free and scaffold‐based cell culture platforms employed to mimic breast tumors and investigate key tumor promoting factors. With special attention to hydrogels within scaffold‐based systems, this review highlights recent advancements driven by artificial intelligence in the design, development, and scalability of hydrogel‐assisted 3D in vitro tumor models, as well as in the detailed investigation of tumor promoting factors. These developments underscore the growing potential of 3D in vitro models as viable alternatives to animal models, influencing translational anticancer strategies, therapeutic innovation, and broader biomedical research.