This study was aimed to investigate the effect of RPMI 1640 and IMDM on the development of human peripheral blood monocyte-derived dendritic cells. Under the same cytokines and culture conditions, the different medium types were tested, and the morphology of mature and immature dendritic cells was observed by microscopy, the cell phenotype and endocytosis ability were detected by flow cytometry. Furthermore, the immunoregulatory function of various DC was analyzed by mixed lymphocyte reaction (MLR), the expression of cytokine in culture supernatant of MLR system was also analyzed by Bio-plex technology. The results showed that there were no difference in morphology, CD14, CD83 expression and endocytosis ability between IMDM-cultured DC and RPMI-1640 medium-cultured DC, but there was a lower expression of CD1a in IMDM-cultured DC. Moreover, DC cultured with IMDM displayed a significant reduction in stimulating T cell proliferation, and highly expressed IL-6, IL-8 and IL-10, but low expressed IL-12. It is concluded that the different cultural mediums can induce DC with different functions and DC cultured with IMDM may correlated with induction of immune tolerance. The results of this study will provide a new idea for DC clinical application.