摘要
目的 探讨体外诱导和培养大鼠骨髓来源树突状细胞（Dendritric cells，DC）的方法，为进一步研究树突状细胞的功能及应用提供理论依据。方法 从健康大鼠管状骨中，无菌下提取骨髓细胞，用GM-CSF和IL-4诱导大鼠骨髓前体细胞，培养10d，在倒置荧光显微镜下观察培养的DC形态学特征，LPS刺激1～2d，进行形态学观察、应用流式细胞仪检测表面分子（未成熟和成熟的）CD80、CD86和OX62的表达水平进行DC表型检测。结果 培养的骨髓细胞胞浆突起大而长，呈树突状，具有DC的典型形态。培养7d未成熟的DC表面共刺激分子CD80、CD86和OX62表型表达的阳性率分别为40．95％、52．93％和45．66％。培养11d的DC表面共刺激分子CD80、CD86和OX62表型表达的阳性率分别为85．93％、81．38％和96．36％。结论 成功建立了大鼠骨髓DCs的体外诱导方法，为研究DC的功能及其在移植耐受中的作用奠定基础。
Objective To explore methods of inducing bone marrow-derivod dendritic cells （DCs） in Dark Agouti （DA） rats and to provide the theory evidence for research of DCs function and application. Methods Bone marrow cells were taken from the hind limbs and co-cultured with cytokine GM-CSF and IL-4. Ten days later, the morphological characters of DC were observed by inversion fluorescence microecope to be treated with LPS for 1 - 2 days. The expressions of CD80 and CD86 were detected by flow cytometry to determine DC phenotype. Results The cultured bone marrow cells exhibited morphological characteristics of DCs, having big and long endochylema ecphyma with dendrite type. Eleven days after culture, the cells had DC phenotype as characterized by 85.93% and 81.38% positive expression rate of the co-stimulating molecule CD80 and CD86. Conclusions The approaches of culturing bone marrow-derived cells with GM-CSF and IL-4 could obtain adequate DCs with high purity leading to success in vitro induction and proliferation of DCs. This method paves the way for research in function of DCs and its action in transplantation toleration.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2007年第3期223-226,共4页
Chinese Journal of Gerontology
基金
教育部留学博士回国科研启动基金资助项目(2004科技No.1173)
教育部博士生科研基金(No.20050183069)
吉林省科技厅科技发展计划资助项目(No.20050408-1)
国家自然科学基金资助项目(No.30670301)
关键词
大鼠
树状突细胞
体外诱导
Dendritic cells
In vitro induction
Rat
