Combining Microwave Ablation With CAR-T-Cell Therapy in Tumor-Bearing Mouse Models.

Microwave ablation (MWA) is a thermal ablation technique widely used for local tumor control that has the added potential to stimulate systemic anti-tumor immunity. Although MWA alone rarely eliminates recurrent or metastatic disease, its ability to remodel the tumor microenvironment makes it a promising partner for adoptive cell therapies such as chimeric antigen receptor (CAR)-T cells. However, reproducible protocols for combining these approaches remain limited. This protocol describes the integration of MWA with CAR-T therapy in tumor-bearing mouse models. Human hepatocellular carcinoma cell lines (Hep3B and SK-HEP-1) are inoculated subcutaneously into NOG mice to establish tumors. Localized MWA is performed at adjustable power and duration to induce partial or complete ablation. At defined intervals following MWA, CAR-T cells derived from healthy donor T cells and transduced with a lentiviral vector are injected intravenously. This experimental design uniquely separates MWA and CAR-T delivery, enabling precise evaluation of thermal preconditioning effects on the tumor microenvironment and subsequent CAR-T activity. By combining localized ablation with adoptive immunotherapy, the protocol provides a translationally relevant platform to optimize treatment timing, enhance CAR-T efficacy in solid tumors, and address key barriers in tumor immunology and cancer therapy. Key features • Practical protocol to investigate MWA and CAR-T-cell combination therapy in mouse tumor models. • Includes detailed procedures for tumor cell preparation, subcutaneous inoculation, MWA treatment, and CAR-T-cell administration. • Novel protocol design separates MWA and CAR-T delivery, allowing rigorous analysis of preconditioning effects on the tumor microenvironment and therapy response. • Offers translational relevance for improving CAR-T therapy in solid tumors, addressing a critical barrier to clinical application.