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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 (DCs) are the cells with the strongest antigen presenting function, but its application is very limited because of its extremely small number. Objective: To establish a large-scale amplification method of mouse bone marrow DC in vitro. Design: Complete randomized controlled study. Location and Subjects: Experimental sites: Central Laboratory of Basic Medicine; 6 C57BL / 6 (H-2kb) mice and 6 BALB / c (H-2kd) mice. Intervention: Mouse bone marrow cells were cultured in vitro with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). MAIN OUTCOME MEASURES: Morphological changes were observed by phase-contrast microscopy. 3H-TdR incorporation assay was used to detect cell proliferation. Cell surface molecules were detected by FACS. Cytokines were detected by ELISA. Results: The morphological characteristics of DCs were typical, showing strong ability to activate T cells and express IaKb, CD11c, CD80, CD86 and ICAM-1 molecules as well as IL-12, IL-6, TNF- and IL- Enhanced level. Conclusion: This method has a lot of DC cells expanded in vitro with strong immunological activity.