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The target tumor cells are labeled with specific fluorescent cytoplasmic reagent, the live cells can be imaged and monitored at real-time level to measure the tumor cell lysis. The reagent is safe for cells at optimized concentration, and fluorescence measurements can be accurately performed for short-term experiments, as the reagent will be diluted with cell proliferation. In addition, the ratio of target tumor cells and CAR-T cells can be optimized in the assay. https://www.creative-biolabs.com/car-t/t-cell-mediated-tumor-cell-lysis-assay.htm
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Reference: https://en.wikipedia.org/wiki/Chimeric_antigen_receptor_T_cell https://www.creative-biolabs.com/car-t/t-cell-mediated-tumor-cell-lysis-assay.htm T Cell-Mediated Tumor Cell Lysis Assay
Background • Chimeric antigen receptors (CARs, also known as chimeric immunoreceptors, chimeric T cell receptors or artificial T cell receptors) are receptor proteins that have been engineered to give T cells the new ability to target a specific protein. The receptors are chimeric because they combine both antigen-binding and T-cell activating functions into a single receptor. • CAR-T cell therapy uses T cells engineered with CARs for cancer therapy. The premise of CAR-T immunotherapy is to modify T cells to recognize cancer cells in order to more effectively target and destroy them. Scientists harvest T cells from people, genetically alter them, then infuse the resulting CAR-T cells into patients to attack their tumors.
Anti-tumor activity in mice is induced only when both the universal CAR T cells plus the correct antigen-specific adaptor molecules are present. Anti-tumor activity and toxicity can be controlled by adjusting the administered adaptor molecule dosing. Treatment of antigenically heterogeneous tumors can be achieved by administration of a mixture of the desired antigen-specific adaptors. Thus, several challenges of current CAR T cell therapies, such as: • the inability to control the rate of cytokine release and tumor lysis • the absence of an “off switch” that can terminate cytotoxic activity when tumor eradication is complete • a requirement to generate a different CAR T cell for each unique tumor antigen • may be solved or mitigated using this approach.
Two Assays for T Cell-Mediated Tumor Cell Lysis • The target tumor cells can be incubated with the radiolabel. The specific radiolabel is taken up and reversibly binds to cytosolic proteins. When the target cells are recognized by CAR-T lymphocytes, the target cells are killed and the radiolabel is released.
Two Assays for T Cell-Mediated Tumor Cell Lysis • The target tumor cells are labeled with specific fluorescent cytoplasmic reagent, the live cells can be imaged and monitored at real-time level to measure the tumor cell lysis. The reagent is safe for cells at optimized concentration, and fluorescence measurements can be accurately performed for short-term experiments, as the reagent will be diluted with cell proliferation. In addition, the ratio of target tumor cells and CAR-T cells can be optimized in the assay.
Reference: https://en.wikipedia.org/wiki/Chimeric_antigen_receptor_T_cell https://www.creative-biolabs.com/car-t/t-cell-mediated-tumor-cell-lysis-assay.htm Thanks