Cultivation and Identification of Dendritic Cells from Mouse Bone Marrow in Vitro

Objective To establish a method of cultivation of dendritic cells(DC) from mouse bone marrow in vitro and identify their phenotype and function. Methods Under aseptic condition,bone marrow cells were extracted from the tibia and femur bones of BALB/c mice. Bone marrow cells were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor(rmGM-CSF) in vitro. The expansion and morphological changes of DC were observed with light inverted microscope. Phenotype was identified with flow cytometry and biological function was studied with antigen phagocytosis test. Results A large number of immature and mature DC with typical dendritic morphological characteristics could be generated from murine bone marrow. Immature DC, which had high expression in CD11c and low expression in CD40, MHC-II and CD86, could phagocytize antigen. Mature DC, which could be induced from immature DC by lipopolysaccharides, had high expression in CD11c, CD40, CD86 and MHC-II molecules. Conclusion Immature and mature DC can be generated from mouse bone marrow cells through cytokine induction in vitro and be used for further study associated with DC.