Culture and identification of mouse bone marrow derived DCs that induce T lymphocyte proliferation

Objective:To establish a method of inducing and culturing dendritic cells from mouse bone marrow in vitro,and observe their biological feature and function of stimulating T lymphocytes proliferate in different stages.Methods:Mononuclear cells isolated from mouse bone marrow were induced into dendritic cells by being cultured with GM-CSF and IL-4,and then examined from aspects of morphology,phenotype and function.Results:Modality of mononuclear cells diversified after being cultured in GM-CSF and IL-4 for 3 days,and growthing clustered-liked.When culturing for 5 days,typical morphology with dendritic processes can be observed.A large number of morphologically typical dendritic cells were observed after culturing for 8 days,under scan electron microscope.DCs displayed typical morphology with dendritic processes and high expressed specific marker of bone marrow derived DC-CD11c and costimulatory molecules of CD40,CD80,CD86 and MHC-Ⅱ.Mature BMDC could stimulate syngenic and allogenic mixed lymphocyte proliferation.Conclusion:A large number of dendritic cells can be generated by culturing the mononuclear cells derived from mouse bone marrow in vitro,and thus will lay a foundation for future research in producing the anti-tumor vaccine.