<h3>Background</h3> Monocyte-derived dendritic cells (MoDCs) represent an important alternative to the challenge of obtaining natural DCs for immunotherapy applications. However, their <i>ex vivo</i> manipulation for MoDCs-based vaccine development poses significant obstacles due to the wide range of antigenic and maturation stimuli alternatives. <h3>Methods</h3> In this study, different antigen formats were compared in combination with three MoDCs maturation protocols to assess their impact on the effector response of CD8+ T cells, using different formats of CMV antigen as a study model (figure 1). <h3>Results</h3> Results showed that the maturation cocktail based on LPS induced the highest expression of maturation markers in MoDCs. Among the different antigen formats, short and synthetic long peptides presented in MoDCs matured with the Standard cocktail (Std-mDCs) promoted the highest cytokine production. Furthermore, CD137 expression positively correlated with the effector cytokine response, especially in the Std culture. LAG-3 expression was observed to be associated with the maturation cocktail used, showing the lowest levels in the Std culture. <h3>Conclusions</h3> These findings suggest that the maturation protocol may impact the effector response of CD8+ T cells by influencing the expression of activating or inhibitory receptors. These results could have significant implications in the design of MoDCs-based vaccines. <h3>Acknowledgements</h3> This project was supported by grants from Minciencias 807 (840-2018), 920 (800-2023), Universidad Nacional de Colombia HERMES 57494, and Fundacion Salud de Los Andes. <h3>Ethics Approval</h3> This project was validated by the ethics committee of the Faculty of Medicine of the National University of Colombia through minute 006 of April 13, 2023.