A patient-derived, autologous component-integrated biomimetic hydrogel system for lung cancer organoid culture and recapitulation of immunotherapy response

Background: Matrigel-based culture of patient-derived lung cancer organoids (LCOs) is limited by undefined composition, batch-to-batch variability, and immunogenicity, which compromise experimental reproducibility and drug response readouts. Methods: We developed an autologous component-integrated hydrogel by incorporating patient-derived decellularized lung fiber fragments (1.25 μm diameter; 52.5 μm length) and optimized autologous plasma (56 °C, 30 min; 8,000×g, 5 min; 5% v/v) into a porous gelatin methacryloyl (GelMA) network. Five formulations—GelMA, GelMA+ fiber, GelMA + plasma, MC-GelMA (fibers + plasma) and Matrigel—were compared to dissect component contributions. LCOs were established from 8 patients, and PBMC co-culture experiments were performed with 3 samples (1 PD-L1-positive, 2 PD-L1-negative). Results: MC-GelMA exhibited uniform porosity and a Young’s modulus of 15.67 ± 0.86 kPa, closely matching native lung tumor tissue. Compared with Matrigel, LCOs cultured in MC-GelMA exhibited in vivo-like morphology, enhanced viability and metabolic activity, and preservation of key histological and genomic features.The platform supported stable long-term culture through passage 9 with maintained drug sensitivity. Component analysis revealed that fibers mainly provided structural and mechanical cues, while plasma supplied biochemical signals; their combination synergistically enhanced organoid growth and immune cell infiltration. In co-culture, MC-GelMA recapitulated anti-PD-L1 responses: PD-L1-positive organoids showed increased LDH release, whereas PD-L1-negative organoids did not respond. Conclusion: In conclusion, this autologous component-integrated biomimetic hydrogel serves as a highly reproducible biofabrication platform for stable LCO culture and reliable recapitulation of patient-specific immunotherapy response patterns. Further validation in larger cohorts is warranted to establish its predictive potential for clinical immunotherapy outcomes.