活体磁共振示踪磁标记大鼠神经干细胞  

In vivo MRI tracking of neural stem cells labeled with Feridex

作　　者：翟宝进[1] 钟士江[1] 王爽[2] 李双英[1] 陈礼明[1] 

机构地区：[1]武警医学院附属医院,天津市300162 [2]哈尔滨医科大学附属医院,黑龙江省哈尔滨市150086

出　　处：《中国组织工程研究与临床康复》2009年第1期61-64,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30370500);天津市自然科学基金面上项目(05YFJMJC14900)~~

摘　　要：背景：要将神经干细胞替代治疗神经系统疾病应用于临床,必须解决一个重要问题,就是植入人脑内神经干细胞的存活、迁移、识别和动态监测。目的：拟建立菲立磁体外标记胎鼠神经干细胞的方法及检测手段,观察标记细胞移植后在活体上磁共振信号的改变。设计、时间及地点：MRI动态评估,体内实验,于2005-01/08在武警医学院附属医院完成。材料：孕13-18dSD胚胎大鼠10只用于分离、培养大鼠胚胎源性神经干细胞,健康成年清洁级SD大鼠32只,用于制作局灶性脑缺血再灌注模型。方法：分离培养大鼠胚胎源性神经干细胞,使用菲立磁-多聚赖氨酸复合物标记神经干细胞。制作局灶性脑缺血再灌注模型,将菲立磁-多聚赖氨酸复合物标记的神经干细胞分别移植入模型大鼠左侧脑内,右侧移植未标记的神经干细胞。主要观察指标：对标记细胞进行普鲁士蓝染色、电镜观察。细胞活体移植后1,5,14d体内磁共振示踪。结果：菲立磁可以高效率地标记神经干细胞,普鲁士蓝染色显示菲立磁-多聚赖氨酸复合物标记神经干细胞胞质内出现细小的蓝色铁颗粒,电镜结果显示菲立磁-多聚赖氨酸复合物标记的神经干细胞胞质内含有许多包裹铁颗粒的囊泡。磁共振成像检查发现脑内移植的标记细胞在磁共振上呈明显的低信号改变,移植第5天低信号物质沿胼胝体腹侧迁移。在移植第14天,对称平行的两个针道已经基本看不见,病灶侧侧脑室部位低信号物质向对侧迁移,左侧侧脑室部位低信号物质基本看不见。结论：菲立磁经多聚左旋赖氨酸转染后可体外标记神经干细胞,标记后体内移植的神经干细胞可以在磁共振上产生明显的低信号改变。BACKGROUND： Monitoring the survival, migration and identification of neural stem cells（NSCs） implanted into the brain is an important problem for its clinical application for nervous system diseases. OBJECTIVE： To evaluate the feasibility of monitoring the neural stem cells implanted into the brain by the technique of labeling with Feridex （FE） and observe nuclear magnetic resonance signal changes. DESIGN, TIME AND SETTING： MRI dynamic evaluation and in vivo trial were performed at the Hospital of Medical College of Chinese People＇s Armed Police Forces between January and August 2005. MATERIALS： Ten SD rats of 13-18 embryonic days were used to isolate and culture embryo-derived neural stem cells （NSCs）; in addition, 32 normal SD rats were used to make middle cerebral artery occlusion （MCAO） model. METHODS： The NSCs were isolated and cultured, and labeled with Feridex mediated by poly-L-lysine （FE-PLL）. The FE-PLL labeled NSCs and normal NSCs were implanted into left and right brain of MCAO model rats, respectively. MAIN OUTCOME MEASURES： The labeled cells were identified by Prussian blue staining and electron microscopy, and the 1, 5, and 14-day images were identified by MRI. RESULTS： Feridex could label the NSCs with high efficiency. Prussian blue staining showed numerous blue-stained iron particles. Transmission electron microscopy revealed the presence of numerous vesicles filled with electron-dense magnetic iron particles in the FE-PLL-labeled NSCs. MRI imaging showed that labeled cells presented obvious low signal on MRI, and the signals migrated along the corpus caliosum on day 5. On the 14{h day, two parallel pin holes had disappeared, and the low signals at the corpus callosum moved to contralateral region. No low signal substance was observed at the left brain. CONCLUSION： FE can be used to label NSCs in vivo, and the labeled NSCs can be tracked in vivo with MRI.

关 键 词：神经干细胞 菲立磁 磁标记细胞 

分 类 号：R394.2[医药卫生—医学遗传学]