Generating Microtumors through 3D Bioprinting of Tumor Organoids: Ink Development and Scalable Manufacturing

Cancer is one of the most challenging global health problems and millions of patients rely on chemotherapy to combat cancer. However, cancer research faces challenges in providing personalized therapeutic regimens tailored to patients’ molecular disease profiles. Developing robust preclinical models for antitumor drug evaluation is critical for new drug discovery and clinical translation. Recent advances in biomedicine integrate 3D culture systems with stem cell-driven self-organization and generated patient-derived tumor organoids (PDTOs). PDTOs preserve intertumoral heterogeneity while exhibiting pharmacological responses closely resembling in vivo drug sensitivity profiles. By leveraging patient-specific tumor biology, these platforms have high transformative potential for precision oncology, enabling both accelerated drug discovery and personalized therapeutic stratification. But conventional tumor organoid culture methods have critical limitations that impede new system development. 3D bioprinting can offer excellent spatial precision and reproducibility, positioning it as an ideal technology for fabricating 3D tumor organoids for biomedical research. In this study, we engineered a functional 3D culture platform for hepatic carcinoma organoids by combining hyaluronic acid (HA, for achieving extracellular matrix mimicry) and gelatin methacryloyl (GelMA, for providing structural fidelity of printed structures) into multi-component bioinks, which exhibited dual-functions of sustaining patient-derived neoplastic organoid propagation and enabling high-fidelity bioprinting. For extrusion 3D bioprinting, the microstructural features, physiochemical characteristics, and rheological behavior of bioinks were systematically studied using different techniques. The influence of critical bioprinting parameters, including extrusion pressure and nozzle moving speed, on structural resolution and fidelity of bioprinted structures was investigated via a parametric analysis framework, employing morphological profiling and filament diameter measurements. We observed the robust expansion of phenotype-stable tumor organoids within the hydrogel matrix, vis-à-vis conventional Matrigel, and established scalable biomanufacturing for generating microtumors containing high-density arrays of monodisperse organoids. The 3D bioprinted organoid-laden microtumors would be employed as high-fidelity platforms for screening chemotherapeutic agents, enabling patient-specific chemotherapy.