Culture and Identification of Dendritic Cells from Human Peripheral Blood in Vitro

Objective To cultivate and identify dendritic cells from human peripheral blood in vitro. Methods The mononuclear cells were obtained from healthy human aseptic fresh peripheral blood by ficoll density gradient centrifugation. The adherent cells were cultured with human recombinant granulocyte-macrophage colony-stimulating factor(rhGM-CSF) and human recombinant interleukin-4(rhIL-4). Tumor necrosis factor-α(TNF-α) was added in 7 days. Then the morphological features were observed by invert optical microscope,we evaluated the phenotypes changes by flow cytometry(FCM). Results Large numbers of dendritic cells could be obtained from human peripheral blood by combination of GM-CSF,IL-4 and TNF-α. Dendritic cells had the typical morphology and phenotypes changes and highly expressed HLA-DR,CD80,CD83,CD86,HLA-DR+ CD83+ and CD80+ CD86+. Conclusion The results indicate that by combination of GM-CSF,IL-4 and TNF-α,large numbers of dendritic cells could be obtained from human peripheral blood. Dendritic cells may typical morphology and phenotypes changes and highly express HLA-DR,CD80,CD83,CD86,HLA-DR+ CD83+ and CD80+ CD86+.