大鼠未成熟树突状细胞体外扩增及功能鉴定

摘要：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。

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

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.