小鼠骨髓树突状细胞体外扩增方法的探索

背景树突状细胞(Dendriticcells,DC)是抗原提呈功能最强的细胞,但因数量极微限制了它的应用.目的建立小鼠骨髓DC体外大量扩增方法.设计完全随机对照研究.地点和对象实验地点基础医学部中心实验室;C57BL/6(H-2kh)小鼠,BALB/c(H-2kd)小鼠各6只.干预用粒-巨噬细胞集落刺激因子(GM-CSF)与白细胞介素4(IL-4)体外培养小鼠骨髓细胞.主要观察指标用相差显微镜观察形态学变化,3H-TdR掺入法检测细胞增殖能力,FACS检测细胞表面分子,ELISA检测细胞因子.结果扩增的DC形态学特征典型,具有强烈的激活T细胞增殖能力,表达IaKb,CD11c,CD80,CD86,ICAM-1分子以及分泌IL-12,IL-6,TNF-α和IL-1β水平增强.结论该法在体外扩增的大量的DC细胞,具很强的免疫学活性.

Approach to in vitro amplification of dendritic cells from murine bone marrow

BACKGROUND: Dendritic cells(DC) act as a powerful antigen presenting cell, but the quantity of DC is quite small which limits its clinical application.OBJECTIVE: To approach a method for in vitro amplifying DC from murine bone marrow cells.DESIGN: A completely randomized control study.SETTING and MATERIALS: Setting: The Central Laboratory of Basic Medicine; C57BL/6(H-2kb) and BALB/c(H - 2kd) rats(6 each).INTERVENTION: Granulocyte-Macrophage colony stimulating factors(GM-CSF) and Interleukin-4(IL-4) were used to in vitro amplify DC from murine bone marrow cells.MAIN OUTCOME MEASURES: Morphological characters of DC were observed by a phase contrast microscope; Cell proliferation was assayed by3H-TdR method; Cell surface molecules were assayed by FACS, and secreted cytokines were assayed by ELISA.RESULTS: Amplified DC showed morphologically typical characters, and showed a strong ability of promoting T cell proliferation, expressed molecules of IaKb, CD11c, CD80, CD86 and ICAM-1, and secreted a higher level of IL-12, IL-6, TNF-α and IL-1β.CONCLUSION: DC, in vitro amplified by this method, shows strong immune activity.