大鼠外周血内皮祖细胞体外分离鉴定及内皮细胞的-第三军医大学学报.DOC

纳米磁性探针转染大鼠外周血内皮祖细胞的磁共振成像实验研究 曹爱红1，2 石红建1 聂芳1 杨芳3滕皋军1 1、210009 南京，东南大学附属中大医院放射科，江苏省分子影像与功能影像实验室；2、2210006 徐州，徐州医学院第二附属医院放射科3、东南大学生物科学与医学工程系 目的:建立稳定的体外分离培养、诱导分化大鼠外周血内皮祖细胞（endothelial progenitor cells，EPCs），及应用纳米磁探针体外标记EPCs的方法，磁共振（MR）进行标记细胞成像。方法: 抽取SD大鼠外周血,应用Ficoll密度梯度离心的方法获得单个核细胞,同时应用VEGF和bFGF 诱导,使之向内皮细胞分化,以免疫细胞化学鉴定贴壁细胞的内皮标志。制备Fe2O3-arginine复合物，对EPCs进行磁探针标记，应用MR进行细胞群成像。结果:经VEGF和bFGF诱导后的EPCs的内皮标志CD31,CD34,Flk21和vWF在不同时段呈阳性表达。普鲁士蓝染色可见铁颗粒位于细胞质内，标记率接近100％；磁共振成像（MRI）显示标记的细胞群信号强度的变化较未标记细胞群信号降低。结论:大鼠外周血中含有EPCs,在体外特殊诱导环境下,可定向分化为内皮样细胞。Fe2O3-arginine可以有效标记EPCs构建纳米生物探针，临床应用型1.5T MR可在体外进行标记细胞群成像。 关键词: 内皮祖细胞;纳米生物探针；内皮细胞;分化；离体；标记；磁共振成像 中图分类号; Magnetic resonance imaging experimental study of nano-bioprobe construction of endothelial progenitor cells which were isolationed from rat peripheral blood labeled with superparamagnetic iron oxide CAO Ai-hong*，SHI Hong-jian，NIE Fang，YANG Fangaojun * Department of Radiology，Zhong Da Hospital，Southeast University，Jiangsu Key Laboratory of Molecular Imaging and Functional Imaging， Nanjing 210009，China Corresponding author：TENG Gao-jun，Email：gjteng@ 【Abstract】 Objective To investigate the isolation , culture , proliferation and differentiation of endothelial progenitor cells ( EPCs) from rat circulating blood and to study of magnetic resonance imaging of nano-bioprobe which wereconstructed with endothelial progenitor cells and superparamagnetic iron oxide(SPIO). Methods The mononuclear cells（MNCs）were isolated f rom SD rat peripheral blood , and EPCselEmail：caooh@163.com 通讯作者：滕皋军，Email：gjteng＠ were selected by adherence method，the expression of cell markers was assessed using immunohistochemistry. EPCs were incubated with Fe2O3-arginine for 24 hours．Intracellular iron was detected by Prussian blue stain． The cells underwent in vitro MR imaging with various sequences. Results The adherent cells stretched and exhibited the clone like morphology after 3d cultivation and proliferated faster then. Immunocytochemistry stain showe