小鼠骨髓未成熟树突状细胞体外扩增及鉴定

In vitro amplification and identification of immature dendritic cells from murine bone marrow

目的 建立体外大量扩增小鼠未成熟树突状细胞 (DC)的方法 ,从形态学、免疫表型和细胞功能试验等方面予以鉴定。 方法 制备小鼠骨髓细胞 ,分别用不同剂量重组小鼠粒细胞巨噬细胞集落刺激因子 (rmGM CSF)培养 ,7d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rmGM CSF培养获得的DC(GMlowDC)具有DC的典型特征 ,细胞表面高表达CD11c,低表达CD4 0、I A/I E ,不表达B7 1,与大剂量rmGM CSF培养获得的DC(GMhighDC)相比 ,其体外刺激未致敏T淋巴细胞增殖的能力较弱。 结论 本实验中获得的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 ,说明所建立的培养未成熟DC的方法是可行的 ;rmGM CSF的剂量与细胞的成熟程度相关 ,一般说来 ,较大剂量的rmGM CSF诱导生成的细胞以成熟DC为主 ,小剂量rmGM CSF诱导生成的细胞以未成熟DC为主。

Objective To establish a method for in vitro amplification of immature dendritic cells from murine bone marrow, and to identify it with morphological, immunological phenotype determination, and functional examination. Methods Dendritic cells from murine bone marrow were cultured with different dosage of rmGM-CSF. The suspending cells were examined with scanning electronic microscope, and the non-sensitized T lymphocyte proliferation was observed by mixed lymphocyte reaction. Results Dendritic cells(DC)cultured in lower dosage of rmGM-CSF(Gm lowDC) exhibited typical characteristics of DCs with high expression of CD11c and low expression of CD40 and I-A/I-E, and non-expression of B7-1 on the surface of the cells. The capacity of Gm lowDC to stimulate the proliferation of non-sensitized T lymphocyte in vitro was weaker than that of Gm highDC. Conclusion Gm lowDC exhibited typical characteristics of DC, immature in cell phenotype and cell functions, suggesting that our methods of immature DCs culturing was feasible. The dosage of rm GM-CSF has direct relationship with the maturation degree of DC. Generally speaking, mature DC was mainly induced by high dosage of rmGM-CSF, while immature DC by low dosage. [