Physically Inactivated Tumor Organoids Enable Rapid and Personalized Enrichment of Cytotoxic T Cells for Solid Tumor Immunotherapy.

Adoptive T cell therapy holds great promise for the treatment of solid tumors but remains constrained by tumor heterogeneity, inefficient neoantigen targeting, and the complexity of T cell manufacturing. Here, we present a patient-specific, broadly applicable platform using physically inactivated tumor organoids (PIOs) to generate tumor-specific cytotoxic T cells ex vivo. Derived from droplet-engineered tumor organoids (DEOs), PIOs preserve the full antigenic repertoire of the patient's tumor without requiring synthetic peptides, antigen-presenting cells, or neoantigen prediction. Using matched tumor tissue and PBMCs from colorectal and liver cancer patients, we show that PIOs activate and expand tumor-specific T cells with enhanced infiltration, selective cytotoxicity, and robust secretion of IFN-γ and IL-2. Multi-round PIO stimulation achieves 80-400-fold expansion of CD8+CD137+ T cells within two weeks. Transcriptomic and epigenetic profiling suggest that PIOs modulate T cell programs linked to migration and persistence. This work redefines tumor organoids as immunotherapeutic materials and establishes a rapid, cost-effective platform for personalized T cell manufacturing. Our findings provide a new translational route for adoptive cell therapy in solid tumors using patient-derived materials.