小鼠骨髓及脾脏来源的树突状细胞培养及鉴定

Culture and Identify Dendritic Cells Originated From Bone Marrow and Spleen of Mouse

目的建立体外培养和扩增树突状细胞(dendritic cell-DC)的方法,为免疫耐受研究提供实验材料和奠定基础。方法在无菌条件下提取ICR小鼠脾脏淋巴细胞和骨髓细胞,以小鼠重组粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素(IL-4)协同诱导下培养,光镜下观察DC的形态,流式细胞仪检测CD80、CD86表达水平。结果骨髓细胞体外诱导培养3天后,光镜下显示细胞表面不规则,呈树突状突起,可见典型的树突状细胞形态,低表达共刺激分子(CD80,CD86)。结论与脾脏细胞相比,骨髓细胞中不仅富含大量的DC的前体细胞而且诱导成DC时间短。

Objective To establish a method of cultivation and purification of dendritic cells（DCs） from mouse bone marrow and spleen in vitro and provide experimental material and establish basis for the study of immunological tolerance.Methods Extracted the ICR mice spleen lymphocytes and bone marrow cells under sterile conditions,and cultured DCs in co-induced of recombinant granulocyte-macrophage colony-stimulating factor（GM-CSF） and interleukin4（IL-4）.Then,the changes of DC morphology were observed under light microscopy and CD80,CD86 expression levels were detected with flow cytometry.Results bone marrow cells appeared irregular and formed dendritic processes after 3 days cultured with stimulating factor in vitro which showed the typical morphology of dendritic cells,and costimulatory molecules（CD80,CD86） were low expression on the cellular suface.Conclusion Compared with the spleen cells,bone marrow cells are not only rich in DC precursor cells but also induced into DCs with a short time.