小鼠骨髓CD117^+造血干细胞定向分化为未成熟树突状细胞及其鉴定  

Directional differentiation of murine CD117^+ hemopoietic stem cells into immature dendritic cells and their identification

作　　者：姜亚卓[1] 田普训[1] 丁小明[1] 李兆伦[1] 管智慧[1] 丁晨光[1] 薛武军[1] 

机构地区：[1]西安交通大学医学院第一附属医院肾移植科,陕西西安710061

出　　处：《南方医科大学学报》2007年第4期450-453,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30371416)~~

摘　　要：目的建立可以在体外稳定地获得大量高纯度未成熟树突状细胞的方法,并对获得细胞进行形态、功能以及表面标志的鉴定。方法取健康C57小鼠骨髓,通过MACS系统分离、纯化CD117^+造血干细胞;使用SCF+IL-3进行体外扩增,在此基础上使用GM-CSF+IL-4+IL-10诱导其定向分化为未成熟树突状细胞;进而在倒置显微镜、扫描电镜以及透射电镜下观察其形态、功能,并使用流式细胞计数法检测其表面标志的表达,对其鉴定。结果SCF+IL-3可以分别在3、5、7d时体外扩增造血干细胞达10.34±1.43、22.65±2.71、54.39±3.08倍;小鼠HSC可被成功诱导分化为未成熟树突状细胞,后者具有吞噬功能,表面树突较为短小,呈毛刺状,表面标志表达情况为CD11c^+、I-A/I-E^(low)、CD40-、CD80-、CD86-。结论使用该方法可以有效地获得大量高纯度的未成熟树突状细胞并对其进行鉴定。Objective To establish a stable method for obtaining large quantity of highly purified immature dendritic cells （imDCs） in vitro, and identify the morphology, function and surface markers of the cells. Methods CD117＋ hemopoietic stem cells （HSCs） were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor （SCF） and interleukin-3 （IL-3）, the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL- 10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry. Results After 3, 5 and 7 days of culture in the presence of SCF＋IL-3, the cells were expanded by 10.34±1.43, 22.65±2.71 and 54.39±3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c＋, I-A/I-E^low, CD40^-, CD80^-, CD86^-. Conclusion The method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.

关 键 词：CD117 造血干细胞 树突状细胞 定向分化 

分 类 号：R392[医药卫生—免疫学]