Amplification in vitro and identification of biological characteristics on dendritic cell from mouse bone marrow

Objective To establish an effective and convenient method in vitro for induction and amplification of dendritic cell (DC) by using mouse bone marrow cell as the precursor cell.Methods The experiment were divided into two groups:GM/4 and GM/4-α.The precursor cell were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor(rmGM-CSF) and interleukin-4(rmIL-4) in vitro,and dendritic cell in the GM/4-αgroup were treated with recombinant mouse tumor necrosis fac-tor -alpha(rmTNF-α) on the fifth day for stimulating forty-eight hours and that in the GM/4 group was used as controls without treatment.The suspending and loosely adherent cell were collected on the fifth and seventh days for examing with scanning electronic microscope and flow cytometry,and their capacity to stimulate allogenetic T cell proliferation was observed by mixed lymphocyte reaction(MLR).Results The two group cell exhibited typical morphological characteristics of DC and had a much higher CDllc expression rate,which was over 75%.Otherwise,GM/4 DC had a lower expression rate of CD40、CD86、MHC-Ⅱ,which was 30.5%、34.2%、45.1%and that in GM/4-αDC was 78.7%、88.3%、96.7%respectively.In MLR,GM/4 DC had the weaker ability for stimulating the proliferation of allogenetic T cell compared with GM/4-αDC.Conclusion The DC could be induced and amplified from mouse bone marrow by using the method,which laid foundation for the further study on the mechanism of DC inducing immunological tolerance after organ transplantation.