Effects of different culture mediums on biological activity and cytotoxicity in vitro induced immune cells

Objective: To investigate the effects of three culture mediums on biological activity such as proliferation,phenotypes,killing effect and cytokine secretion.Methods: Peripheral blood mononuclear cells(PBMC) were cultured with C3McAb,IL-2 and IFN-γ in serum-free medium MD-CM-STM-N or AIMV or standard serum-containing RPMI 1640 medium separately.Proliferation,phenotypes and cytokine secretion of cells cultured in the three different medium were compared.The cellular proliferative capacity was evaluated by Trypan Blue living cell count,and so did efficiency in killing different tumor cells by MTT,the expression level of the secretory IFN-γ by ELISA,and phenotypes by flow cytometer(FCM).Results: Compared to cells cultured in standard serum-containing RPMI 1640 medium,the proliferation peaks of cells cultured in serum-free medium MD-CM-STM-N and AIMV were prolonged to the 13th day,and in the serum-free medium MD-CM-STM-N group ability of proliferation and the killing activity were significantly increased(P0.05).More percentage of cells cultured in serum-free medium expressed CD8+,CD3+and CD56+(P0.05),cells cultured in serum-free medium secreted more IFN-γ.Conclusion: The proliferative capacity,expression rate of phenotypes,cytotoxicity and cytokine secretion level of immune cells in the serum-free medium MD-CM-STM-N and AIMV are higher than those of them in standard serum-containing RPMI 1640 medium,and MD-CM-STM-N has the advantage of proliferative capacity which provides the experimental and theoretical basis for cytokine-induced immune cells therapy.