Magnetic Nanoparticle-Induced Cell 3D Magnetic Levitation Culture and Drug Evaluation Study

Traditional 2D cell culture systems fail to accurately replicate the intricate tumor microenvironment in vivo, highlighting their growing limitations in preclinical research. In contrast, 3D cell culture technologies have emerged as a prominent research focus due to their capacity to mimic in vivo cellular growth conditions and physiological functionality. While conventional 3D culture systems have improved tumor biology research, they face critical limitations in scalability, reproducibility, and operational complexity. To address these challenges, we developed a magnetic levitation-based 3D cell culture platform utilizing Fe 3 O 4 @DA-PEG@PLL nanoparticles, which enhance cellular adhesion and enable scalable 3D monoculture or multicellular coculture. This method generates uniform three-dimensional cell aggregates (3DCAs) characterized by proliferative capacity, high viability, and minimal interaggregate variability, demonstrating exceptional homogeneity and reproducibility suitable for large-scale cell experiments. Drug resistance assays performed on 4T1 breast cancer-derived 3DCAs and 4T1/3T3 fibroblast coculture models revealed significantly enhanced resistance to clinical drugs compared to 2D cultures. Comprehensive characterization confirmed the ability of the platform to reconstruct tumor microenvironmental features. Protein expression and transcriptomic analyses further elucidated the regulatory effects of 3D culture conditions on protein expression and gene regulation in 4T1 cells.