BALB/c小鼠骨髓来源高产量成熟树突状细胞的制备和初步鉴定  

Preparation and Identification of Dendritic Cells Derived from Bone Marrow Cells of BALB/c Mouse

作　　者：谢峻[1] 王军[2] 郑颖城[3] 董玉梅[3] 王琼书[3] 

机构地区：[1]广州军区武汉总医院科训科,湖北武汉430070 [2]广州军区武汉总医院血液科,湖北武汉430070 [3]广州军区武汉总医院感染控制科,湖北武汉430070

出　　处：《华南国防医学杂志》2014年第6期524-527,共4页Military Medical Journal of South China

摘　　要：目的研究制备、鉴定小鼠骨髓来源的高产量树突状细胞(dendritic cells,DC)的方法,为制备DC及体外研究提供依据。方法取BALB/c小鼠骨髓细胞,经重组小鼠巨噬细胞-粒细胞集落刺激因子(recombinant mouse granulocyte-macrophage colony stimulating factor,rmGM-CSF)、重组小鼠白细胞介素4(recomtinant mouse interleukin 4,rmIL-4)诱导、脂多糖(lipopolysacchrides,LPS)刺激制备高纯度DC。利用光学显微镜、扫描电镜对所制备细胞做形态学观察;流式细胞术检测特定表型标记分子表达情况;酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、干扰素γ(interferon gamma,IFN-γ)的分泌量。从形态、表型标记、细胞因子分泌功能三方面对所制备细胞进行初步鉴定。结果所制备细胞具有典型的DC形态学特征,细胞表面高表达CD1a、CD11c、CD80和CD86,相对于普通DC细胞有大量的TNF-α和IFN-γ分泌。结论利用BALB/c小鼠骨髓细胞在细胞因子rmGM-CSF、rmIL-4诱导及LPS的刺激下成功制备出纯度较高的成熟DC。Objective To study the preparation and identification methods of dendritic cells（IX;s）from bone mar- row cells of mice, and supply reference for the cell preparation and in vitro research.Methods The bone marrow cells from BALB/c mice were treated with GM-CSF, IL-4 and LPS to prepare matured I）Cs.Optical microscopy and scanning electron microscopy were applied to observe the morphological character of the prepared cells. The markers of IX2s on the cells＇ surface and some cytokines secreted by the cells were detected.Results Typical morphological characters were observed in the cells.The expressions of CDla, CD] lc, CDS0, and CD86 in the cells and also the secretion of TNF-a, IFN-y were in creased significantly.Conclusion The treatment with GM-CSF,IL-4, LPS to BALB/c mouse bone marrow cells can induce high production of mature DCs in vitro.

关 键 词：树突状细胞 制备 鉴定 标记分子 细胞因子 

分 类 号：R446.1[医药卫生—诊断学]