大鼠骨髓来源树突状细胞的体外诱导扩增及其功能鉴定

目的探讨体外诱导和扩增大鼠骨髓来源树突状细胞（DC）的方法，并观察其表型及功能特性。方法取得大鼠骨髓细胞后，去除红细胞，加入重组大鼠粒细胞／巨噬细胞集落刺激因子（rrGM-CSF）、重组大鼠白细胞介素4（rrIL-4）培养2周；在光镜和扫描电镜下观察培养DC的形态学特征，流式细胞仪检测DC表面MHC-Ⅱ、CD80、CD86的表达水平，混合淋巴细胞反应检测其刺激同种T细胞增殖能力。结果细胞培养8d后，经倒置显微镜和电镜观察DC出现典型形态，培养6d的DC具有不成熟表型，流式细胞仪检测MHC-Ⅱ为（29．03±4．39）％、CD80为（21．98±7．08）％，CD86为（25．94±6．80）％，刺激同种T细胞增殖能力极低，而培养12dDC的MHC-Ⅱ为（74．05±5．97）％、CD80为（79．85±6．53）％，CD86为（81．00±7．47）％，刺激同种T细胞增殖能力明显增强。结论大鼠骨髓来源干细胞可经联合应用细胞因子诱导培养为具有典型形态特征及功能的DC，为进一步研究其在移植免疫学中的应用奠定基础。

In vitro induction,proliferation and function characterization of dendritic cells from rat bone lnRITOW cells

Objective To establish a method of inducing dendritic cells(DC)from rat bone marrow cells in vitro,and identify the phenotype and function characteristics.Methods The rat bone malToW cells were collected and cultured in vitro under the condition of recombinant rat GM-CSF(rrGM-CSF)and recombinant rat IL-4(rrIL-4).After 2 weeks,the morphological character of DCs was observed under light microscope and scanning electron microscope.Expression of MHC-Ⅱ,CD80 and CD86 were detected by flow cytometry.The ability to stimulate allogenic T cells of the cultured DCs was detected by mixed lymphocyte reaction.Results DCs showed typical morphology with elongated dendritic processes under inversion microscope and scanning electron microscope.DCs at day 6 revealed immature phenotype,including MHC-Ⅱ(29.03 ±4.39)%,CD80(21.98±7.08)%and CD86(25.94±6.80)%.DCs at day 12 showed higher expression of MHC-Ⅱ(74.05±5.97)%,CD80(79.85±6.53)%and CD86(81.00±7.47)%,and stimulatory capacity of allogenic T cells,compared with that in DCs at day 6.Conclusion Matured DCs could be generated from rat bone marrow cells and attendance with rrGM-CSF and rrIL-4,which present the feasibility for further research on its application to allograft immunorejection.