大鼠未成熟树突状细胞体外扩增及功能鉴定
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王为忠

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Amplification and functional identification of immature dendritic cells from rat bone marrow in vitro
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    摘要:

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    目的 探讨建立大鼠体外大量扩增未成熟树突状细胞(DC) 的方法, 以及不同剂量粒细胞巨噬细胞集落刺激因子(GMCSF)对大鼠DC分化成熟的影响。
    方法 分离纯化并扩增大鼠骨髓细胞,用不同剂量GMCSF培养,6 d 和10 d后收集悬浮细胞进行扫描电镜观察和免疫表型鉴定,并行混合淋巴细胞反应,观察其诱导未致敏T 淋巴细胞增殖的情况。
    结果 小剂量GMCSF 培养获得的DC(GMlowDC) 形态上具有DC 的典型特征,在细胞表型、细胞功能试验上具有未成熟的特性,具有DC 的典型特征,细胞表面高表达CD11c,低表达CD80,CD86及MHC II类分子,与大剂量GMCSF加IL4的联合组培养获得的DC( GMhighDC) 相比,其体外刺激未致敏T 淋巴细胞的增殖能力较弱.
    结论 笔者所建立的培养未成熟DC 的方法是可行的;GMCSF的剂量与细胞的成熟程度相关。

    Abstract:

    Abstract:Objective To establish a method for amplification of immature dendritic cells(DC) from murine bone marrow in vitro and investigate correlations between maturation degree of DC and varying dosages of granulocytemacrophagecolony stimulating factor (GMCSF).
    Methods Dendritic cells from murine bone marrow were cultured with different dosages of rm GMCSF. The suspension cells were examined with scanning electronic microscope,and the nonsensitized T lymphocyte proliferation was observed by mixed lymphocyte reaction.
    Results DC cultured in lower dosage of rmGMCSF (GMlow DC) exhibited typical characteristics of DC, and had immature characteristics in cell phenotype and cell functions with high expression of CD11c and low expression of CD80,CD86 and MHC II on the surface of the cells.The ability of GmlowDC to stimulate the proliferation of nonsensitized T lymphocyte in vitro was weaker than that of GmhighDC.
    Conclusions The methods of immature DCs culturing establised by allthors was feasible. The dosage of rm GMCSF has a direct relationship with the maturation degree of DC.

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李纪鹏, 黄怡, 王为忠, 董光龙, 杜建军 ,陈冬利, 季刚, 刘骥.大鼠未成熟树突状细胞体外扩增及功能鉴定[J].中国普通外科杂志,2007,16(9):9-.
DOI:10.7659/j. issn.1005-6947.2007.09.009

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  • 收稿日期:2007-03-20
  • 最后修改日期:2007-09-05
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  • 在线发布日期: 2007-09-25