Isolation,culture,and immunological character of dendritic cells derived from rat bone marrow

Objective To explore the methods for isolating and culturing dendritic cells derived from rat bone marrow in vitro,and provide reference for immunological research and clinical application.Methods The bone marrow mononuclear cells(BMMNC) were obtained from bone marrow of Fischer344 rat's limbs through density gradient centrifugation;and the mature DC was gained by BMMNC cultured in medium with recombinant rat granulocyte-macrophage colony stimulating factor(rrGMCSF),recombinant rat interleukin-4(rrIL-4) and recombinant rat tumor necrosis factor-α(rrTNF-α).In group 1,the rat's ovarian cancer cell line NuTu-19 freeze-melt antigens were added on the third day and the tumor cells lysates pulsed DC was acquired;in group 2,as control,nothing was added.Then the shape of DC was observed,the cell-surface phenotype and the proliferation rate were detected.Results The mature DC could be induced by BMMNC co-culturing with relative cytokines,and the expressions of cell-surface phenotypes such as OX62,CD86 and MHC-Ⅱ in group 1 were higther than those in control group(P0.01).When the ratios of stimulating cells(SC) to reaction cells(RC) were 1∶3,1∶10,1∶30 and 1∶100,the stimulating indexes(SI) in group 1 were higher than those in control group(P0.01).Conclusion The culture method of DC from bone marrow of Fischer344 rats is established,which means the mature DC can be acquired from BMMNC.