大鼠骨髓树突状细胞的体外分离培养及生物学特性研究  

Study on Isolation and Biology of Rat Bone Marrow-Derived Dendritic Cell in vitro

作　　者：叶建新[1] 陈卫昌[1] 黄小平[2] 高楠[1] 

机构地区：[1]苏州大学附属第一医院消化科,江苏苏州215006 [2]苏州大学附属第一医院感染科,江苏苏州215006

出　　处：《苏州大学学报（医学版）》2008年第3期357-359,F0002,共4页Suzhou University Journal of Medical Science

基　　金：江苏省135重点医学人才基金资助项目(RC2007076)

摘　　要：目的探讨体外分离培养大鼠骨髓来源树突状细胞(DC)的方法并对其生物学特性进行研究。方法应用粒-巨噬细胞集落刺激因子(GM-CSF)和IL-4诱导培养从大鼠管状骨中提取的骨髓细胞5 d后,将细胞分成脂多糖(LPS)刺激组和未刺激组(对照组),48 h后进行形态学观察,用流式细胞仪检测组织相容性复合体Ⅱ(MHC-Ⅱ)和CD80的表达,用ELISA法测定培养上清中IL-6和IL-12的含量。结果培养的大鼠骨髓细胞具有树突状突起,具有DC的典型形态。培养7 d后,对照组MHC-Ⅱ、CD80表型表达阳性率分别为57.3%和29.1%,LPS刺激组MHC-Ⅱ、CD80表型表达阳性率分别为70.9%和73.5%;LPS刺激组IL-6和IL-12含量高于未刺激组,差异有统计学意义(P<0.05)。结论成功建立了大鼠DC体外分离培养和诱导成熟的方法;在分化成熟过程中,DC在表型和细胞因子分泌功能上均发生了变化。Objective To explore methods for isolation of rat bone marrow-derived dendritic cell （DC） and to investigate its bionomics. Methods Bone marrow cells were taken from the hind limb and co-cultivated with cytokine GM-CSF and IL-4 for 5 days. The morphological characters of DC were observed by a inversed microscope, the cells were stimulated for 48 hours then morpho- logical characters were observed again. Its MHC-Ⅱ and CD80 molecules were detected with FCM, its IL-6 and IL-12 in co-culture medium was quantified by ELISA. Results Cultivated DC derived from bone-marrow cells have ecphyma and morphological characteristics of DC. MHC-Ⅱ and CD80 phenotype were characterized by 57.3% and 29.1% positive expression rate on untreated DC 7 days later,and 70.9% and 73.5% on DC treated with LPS. The quantity of IL-6 and IL-12 in LPS stimulated culture medium was higher than unstimulated （P〈0.01）. Conclusion A approach of DC derived from bone-marrow cell is established, DC change phenotype and secrete cytokine are discovered maturation in vitro.

关 键 词：大鼠 树突状细胞 分离培养 生物学特性 

分 类 号：R392.11[医药卫生—免疫学]