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超顺磁性氧化铁纳米粒子标记人骨髓和脐带间充质干细胞
作者姓名:马 艳  张德清  陈 乐  王 俭  张 雪  侯 燕  毕晓娟  杨 蓉  胡安华
作者单位:新疆医科大学临床医学研究院,1干细胞研究室,3流式细胞室,新疆维吾尔自治区乌鲁木齐市 830054;新疆医科大学第一附属医院,2影像中心,4产科,5输血科,新疆维吾尔自治区乌鲁木齐市 830054
摘    要:背景:优化人源细胞安全、有效的标记参数及细胞移植后的定位是评价其治疗效果至关重要的环节。 目的:比较核磁影像对比剂超顺磁性氧化铁纳米粒子体外标记人骨髓和脐带间充质干细胞的细胞活率、标记效率及核磁共振T2*WI成像的效果,优化标记细胞处理细节。 方法:培养第3代人骨髓和脐带间充质干细胞,以5-30 mg/L菲立磁(Feridex Ⅳ)结合硫酸鱼精蛋白标记细胞。 结果与结论:人骨髓和脐带间充质干细胞标记前后细胞的存活率接近(P > 0.05)。以5-30 mg/L菲立磁标记骨髓间充质干细胞的阳性标记率差异无显著性意义(P > 0.05);而5 mg/L菲立磁标记脐带间充质干细胞的阳性标记率与20和30 mg/L菲立磁标记的差异有显著性意义(P < 0.05);10 mg/L菲立磁标记脐带间充质干细胞的阳性标记率低于骨髓间充质干细胞(P < 0.05)。当≥20 mg/L菲立磁标记细胞时,2种来源的细胞悬液中均出现不易洗脱和过滤去除的氧化铁颗粒。标记后细胞在3.0T MR GRE T2*WI扫描均见随菲立磁的浓度升高,信号强度减弱。提示这2种组织来源的间充质干细胞,以10 mg/L的菲立磁-硫酸鱼精蛋白复合物标记是安全有效的,可用临床T2* WI的MR成像观察。

关 键 词:核磁影像对比剂  超顺磁性氧化铁  菲立磁  标记  过滤  骨髓间充质干细胞  脐带间充质干细胞  硫酸鱼精蛋白\细胞移植\干细胞  组织工程  
收稿时间:2012-07-17

Superparamagnetic iron oxide nanoparticles label human bone marrow and umbilical cord mesenchymal stem cells
Authors:Ma Yan  Zhang De-qing  Chen Le  Wang Jian  Zhang Xue  Hou Yan  Bi Xiao-juan  Yang Rong  Hu An-hua
Institution:1Laboratory of Stem Cell Research, 3Flow Cytometry Laboratory, Clinical Medicine Research Institute of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Imaging Center, 4Department of Obstetrics, 5Department of Blood Transfusion, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
Abstract:BACKGROUND:Nowadays, it is becoming more and more important to optimize safety of human derived cells, label cells efficiently and track cells after cells transplantation both in basic research and clinic application. OBJECTIVE:To compare the cell viability, labeling efficiency and imaging effect of the T2* weight image (WI) magnetic resonance (MR) between the human bone marrow and umbilical cord derived mesenchymal stem cells labeled with the superparamaganetic iron oxide nanoparticles, as well as to optimize their treatment efficiency. METHODS:The third generation of human bone marrow and umbilical cord derived mesenchymal stem cells were cultured, and labeled with 5-30 mg/L Feridex Ⅳ and protamine sulfate. RESULTS AND CONCLUSION:The viability of human bone marrow mesenchymal stromal cells was similar with human umbilical cord derived mesenchymal stem cells (P > 0.05). There was no significant difference of labeling rate between the bone marrow msenchymal stem cells labeled with 5-30 mg/L Feridex Ⅳ (P > 0.05); while there was significant difference of labeling rate between the umbilical cord derived mesenchymal stem cells labeled with 5 mg/L Feridex Ⅳ and 20 and 30 mg/L Feridex Ⅳ (P < 0.05); the positive labeling rate of umbilical cord derived mesenchymal stem cells was lower than that of bone marrow msenchymal stem cells after labeled with 10 mg/L Feridex Ⅳ (P < 0.05). When two sources of cells were labeled with Feridex Ⅳ more than 2 mg/L, the iron oxide particles were found in the cell suspension and could not be removed by elution and filtration. The signal intensity from 3.0T MR GRE T2*WI scan was decreased with the increasing of Feridex Ⅳ concentration in both cell types. It is safe and effective to label the two tissue-derived mesenchymal stem cells with 10 mg/L Feridex Ⅳ-protamine sulfate complex, and can be observed with T2*WI MR.
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