Generation and Expansion of MART-1 Antigen-Specific T Cells for Immunological Assays

This research aimed to investigate whether T cells could be engineered to recognize and eliminate cancer cells expressing the MART-1 antigen, commonly found on melanoma cells. The goal was to inform strategies for improving cancer immunotherapies by generating and expanding antigen-specific T cells in vitro. Donor T cells were thawed and transfected with MART-1 RNA to induce expression of the antigen-specific receptor. Post-transfection, Fluorescence-Activated Cell Sorting (FACS) confirmed receptor expression, and the cytotoxic activity of modified T cells was evaluated using the Incucyte live-cell imaging system. Transfection efficiency varied between batches, with 7.4% in one and 36.8% in another, resulting in limited cancer cell killing. Factors contributing to these results may include low RNA quality, poor post-thaw cell recovery, or variability in the initial donor cells. Although the experiment did not achieve full cytotoxic functionality, it highlighted key technical challenges and provided insights for optimizing T cell engineering protocols. These findings contribute to understanding how to generate and expand functional, antigen-specific T cells for potential therapeutic applications in cancer immunotherapy.