Three-dimensional cell culture models in oncology: from tumour spheroids to integrated microphysiological systems

Traditional two-dimensional (2D) cell cultures fail to capture the structural, mechanical, and biochemical complexity of living tissues, prompting a major paradigm shift toward three-dimensional (3D) cell culture models. 3D systems faithfully replicate in vivo physiological architectures, dynamic cell-to-cell contacts, and extracellular matrix (ECM) interactions, bridging a critical gap in preclinical studies by predicting human therapeutic responses more accurately. This review explores the technological evolution from simple multicellular tumor spheroids to sophisticated microphysiological networks. Scaffold-free methods-including the hanging drop, low-adherent plates, magnetic levitation, and dynamic bioreactors-efficiently recreate the pathophysiological oxygen, nutrient, and metabolic gradients characteristic of solid tumor zonation and drug-resistance pathways. Conversely, scaffold-based systems leverage hydrogels, synthetic polymers, and decellularized matrices to dissect the biomechanics of matrix stiffness and mechanotransduction. At a higher tier of biological fidelity, patient-derived organoids preserve cellular heterogeneity, native tissue polarity, and accurate patient-specific mutational profiles, serving as critical pipelines for personalized medicine and precision drug screening. Furthermore, the integration of advanced microfluidic technologies has facilitated the engineering of tumor-, organ-, and multi-organ body-on-a-chip platforms. These microphysiological systems reconstruct dynamic fluid flow, functional tissue-vascular interfaces, physiological barriers (such as the blood-brain barrier), and systemic pharmacokinetic/pharmacodynamic (ADME) drug profiles. Beyond deepening mechanistic insights into oncology, these interconnected 3D models provide highly ethical, predictive, and cost-effective alternatives to animal experimentation in strict alignment with the 3Rs principles. Ultimately, these complementary 3D culture technologies are transforming oncology into a highly predictive, patient-tailored discipline.