摘要
目的:探讨经MACS系统在体外获得大量高纯度未成熟树突状细胞(imDC)的方法,并对其细胞表面标志、形态和功能进行鉴定。方法:对健康C57小鼠的骨髓通过MACS系统分离、纯化CD117+造血干细胞(HSC);使用SCF+IL-3行体外扩增,并采用GM-CSF+IL-4+IL-10诱导其定向分化为imDC;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,采用流式细胞计数法检测、鉴定其表面标志物的表达。结果:SCF+IL-3可以分别在3、5、7d时体外扩增HSC达10.34±1.43倍、22.65±2.71倍、54.39±3.08倍;小鼠HSC可被成功诱导分化为imDC,且具有吞噬功能,表面树突呈毛刺状、较为短小,imDC并表达CD11c+、I-A/I-Elow、CD40-、CD80-、CD86-。结论:本方法可稳定、有效地获得大量高纯度的imDC。
AIM： To Investigate a method that can obtain massive highly purified immature dendritic cells（imDCs） steadily in vitro, and identify them by morphous, function and surface markers by MACS. METHODS： Isolate and purify CDl17^＋ hemopoietic stem cells （HSCs） from bone marrow of healthy C57 murine by MACS. After being expanded by SCF ＋ IL-3, HSCs would be directional differentiated into imDCs by use of cytokine scheme of GM-CSF ＋ IL-4 ＋ IL-10. Then identify imDCs through following ways： observing morphous and function of them under inverted microscope, scanning electron microscope and transmission electron microscope, detecting the expression of surface markers by flow cytometry. RFSULTS： The fold of expansion 3, 5 and 7 days after cultured with SCF ＋ IL-3 was separately 10.34 ＋ 1.43, 22.65 ＋ 2.71 and 54.39 ＋ 3.08. HSCs can be successfully differentiated into imDCs, which have the function of phagocytosis. The imDCs were short and small, in shape of sentus. The expression of surfacee markers was CD11c^＋ , I-A/I-E^low, CD40^-, CD80^-, CD86^- by flow cytometry. CONCLUSION： This method can obtain and identify massive highly purified imDCs steadily in vitro.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2007年第10期917-920,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(30371416)
