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

ABSTRACT 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.