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目的建立体外诱导和扩增大鼠骨髓树突状细胞(DCs)的方法,并进行生物学鉴定。方法大鼠骨髓细胞经贴壁去除悬浮细胞,加入细胞因子(IL-4和GM-CSF)培养2周。经过光镜,透射电镜观察培养DCs的形态学特征;通过同种T细胞混合培养,采用MTT比色分析法测定不同浓度的DCs发同种T细胞增殖的能力。结果培养的DC胞浆突起大而长,呈树突状,具有DCs的典型形态,并且具有强烈的激发同种异体T细胞增殖的能力。结论采用大鼠骨髓细胞经贴壁去除悬浮细胞,加入细胞因子(IL-4和GM-CSF)序贯培养,能获得大量、较高纯度的DCs。
OBJECTIVE: To establish a method for inducing and amplifying rat bone marrow dendritic cells (DCs) in vitro and to conduct biological identification. Methods Bone marrow cells were removed by adherent cells and cultured for 2 weeks with addition of cytokines (IL-4 and GM-CSF). Morphological characteristics of cultured DCs were observed under light microscope and transmission electron microscope. The proliferation of DCs stimulated by different concentrations of DCs was measured by MTT colorimetric assay by mixed culture of allogeneic T cells. Results The cultured DCs had large and long protuberances, which were dendritic in shape. They had the typical morphology of DCs and had strong ability of stimulating the proliferation of allogeneic T cells. CONCLUSIONS: Adherent cells of rat bone marrow cells are used to remove suspended cells and cytokines (IL-4 and GM-CSF) are sequentially cultured to obtain a large amount of DCs with high purity.