Triple-negative breast cancer (TNBC) represents a significant clinical challenge due to its aggressive nature and lack of targeted therapies. Recent studies highlight a correlation between glucose metabolism and tumor progression, with emerging evidence suggesting that glucose restriction may serve as an effective adjuvant therapeutic strategy. To address the limitations of traditional in vitro and in vivo experimental models adopted for translational research and drug discovery, an advanced 3D in vitro TNBC model was developed utilizing MDA-MB-231 cells embedded in tunable stiffness alginate-based hydrogels integrated into an organ-on-chip platform (MIVO®) resembling dynamic culture conditions that better mimic the in vivo-like diffusion of nutrients, drugs, and biochemical signals.