Production of functional dendritic cells from mouse bone marrow

Currently, immune cell-based therapies, particularly those that utilize dendritic cells (DCs), are a promising therapy approach for cancer treatment. Therefore, DC therapy is the focus of many studies in many laboratories worldwide that are developing novel cancer therapies. This study aimed to develop a reproducible procedure to produce functional DCs from mouse bone marrow for DC therapy research. Bone marrow was collected from mouse femur bones by flushing with buffered saline. These cells were used to isolate mononuclear cells (MNCs) by Ficoll gradient centrifugation. MNCs were cultured in RPMI 1640 medium supplemented with 20 ng/mL of IL-4 and 20 ng/mL of GMCSF to induce maturation of immature DCs. The results showed that this procedure induced cells exhibiting the DC phenotypes, such as the expression of CD40, CD80, and CD86, high phagocytic capacity, strong production of IL-12, and efficient stimulation of T-CD4 lymphocytes. These results suggest that this procedure can be used to produce functional DCs in future studies that use DCs for immune therapy.