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Introduction: Dendritic cells (DCs) play crucial roles in cellular immunity as the most powerful antigen presenting cells. They have been widely used for antigen delivery in vivo and in vitro. There are different ways to generate DCs and also gene transduction. In this study we introduce some optimization in order to produce high amount of well differentiated murine DCs for potential immunotherapy and vaccine development applications. Methods: Murine bone marrow cells were isolated from male BALB/c mice and the cells were cultured with complete RPMI in presence of the same ratio of IL-4 and GM-CSF. Some changes were made in the medium and the lysis buffer applications to increase the differentiation rate of the cells. Lentiviral virions were applied to transfer the genes of interest to DCs with no pre-maturation steps. CD11c, MHC-II, CD80 and CD86 were assessed by flow cytometry. Results: The optimized steps led to significant increase in number of the isolated cells. IL-4 usage in a similar dose to GM-CSF led to macrophage formation inhibition. Lentiviral virions resulted in successful gene delivery along with well-maturated DCs. Conclusion: The introduced optimized steps could be followed in different DC applications by using lentiviral virions to transduce DCs, independent of the pre-maturation steps.

Keywords: Dendritic cells, Murine, Optimization, Transduction, Lentiviral vectors.

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