Adoptive immunotherapy with CMV-specific T cells (CMV-VSTs) has shown favorable efficacy and minimal adverse effects in clinical settings, serving as prophylaxis, preemptive and/or curative treatment for the restoration of CMV-specific immunity in patients after allogeneic hematopoietic stem cell transplantation. The establishment of a CMV-VST bank enables the prompt use of CMV-VSTs as off-the-shelf therapeutics. CMV-VSTs can be generated through cell culture or immunomagnetic selection based on IFN-γ secretion or multimer technology. In this study, we investigated the feasibility of generating CMV-VSTs via cell expansion after IFN-γ based immunomagnetic isolation, with the goal of establishing a good-quality, cost-effective local production approach. We also assessed the value of incorporating transcriptomic analysis into the current T cell evaluation framework. Our results demonstrate that good-quality CMV-VSTs can be produced using either autologous feeder cells or feeder cells from the rapid expansion protocol (REP), in combination with cytokines such as IL-2 or IL-7/IL-15. Phenotypic and functional analyses confirmed the consistent quality of the final T cell products and showed no significant differences across the various combinations of feeder cells and cytokines. However, transcriptomic analysis highlighted the advantages of using IL-7/IL-15 and autologous feeder cells. Collectively, our findings offer new insights into future strategies for the manufacturing of antigen-specific T cells and underscore the importance of comprehensive, multidimensional assessment in T cell-based immunotherapies.