小鼠骨髓树突状细胞体外扩增方法的探索(英文)

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

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-2kb)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。

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 by 3H 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 αand IL 1β. CONCLUSION: DC, in vitro amplified by this method,shows strong immune activity.