Methods of CD103+ dendritic cells differentiation from bone marrow

Objective:This study was performed to explore methods of inducing and culturing CD103+ dendritic cells(DCs) from mouse bone marrow mononuclear in vitro.Methods:The bone marrow mononuclear cells were obtained from femurs and tibias,and purified by mononuclear cells separation solution.Cells incubated with granulocyte-macrophage colony-stimulating factor (GM-CSF)and Fms-like tyrosine kinase-3 ligand (FLT3L)were added to induce cell differentiation.The morphology,phenotype and distribution were observed under invert microscope.The expression of CD103 + DCs and co-stimulatory molecules were detected by flow cytometry.CD103+ DCs were sorted by magnetic beads and detected the purity.Results:After 8 days culture,the mononuclear cells were differentiated into various forms under the microscope.Most of them were round or irregular,various morphologies and distribute clustered with plenty of dendrites.The expression of CD11c,CD103 and co-stimulatory molecules elevated significantly,and the purity of CD103 + DCs was 95.7％.Besides,the CD103 + DCs could stimulate T cell differentiation efficiently.Conclusion:CD103+ DCs can be efficiently induced by GM-CSF and FLT3L in vitro.