Isolation, culture, induction and identification of different subsets of human dendritic cells

Objective To isolate, culture and induce different subsets of dendritic cells from human peripheral blood mononuclear cells (PBMC). Methods PBMCs were prepared from buffy coats of peripheral blood by centrifugation through a Ficoll density gradient. Then dendritic cells were isolated using the specific monoclonal antibody magnetic microbeads. After adding different cytokines, cells were differentiated into different stages. Results The purity of dendritic cells was greater than 95% as assessed by flow cytometry. Mature denendritic cells expressed higher levels of HLA-DR, CD86, CD40 than immature dendritic cells. There were also morphological differences between two subsets of dendritic cells. Conclusion We have developed the methodology for isolating, culturing and inducing the dendritic cells of different stages in vitro.