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浙江大学学报(医学版)  2017, Vol. 46 Issue (5): 481-486    DOI: 10.3785/j.issn.1008-9292.2017.10.05
精准影像医学专题     
Molday IONTM EverGreen标记大鼠骨髓内皮祖细胞及体外磁共振成像研究
杨荣, 尧林鹏, 张思影, 陈峰
浙江大学医学院附属第一医院放射科, 浙江 杭州 310003
Rat endothelial progenitor cells labeled with Molday IONTM EverGreen and in vitro MRI study
YANG Rong, YAO Linpeng, ZHANG Siying, CHEN Feng
Department of Radiology, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China
 全文: PDF(4168 KB)  
摘要:

目的:探讨用双模态对比剂Molday IONTM EverGreen (MIEG)标记大鼠骨髓来源内皮祖细胞(EPC)及其行磁共振扫描的可行性。方法:以铁的终浓度为10、20、50 μg/mL的MIEG对大鼠骨髓来源EPC进行磁性和荧光双标记,普鲁士蓝染色及荧光镜下观察细胞标记情况,比较不同浓度MIEG标记EPC后的标记阳性率;台盼蓝染色检测20 μg/mL MIEG标记EPC后1 d及1、2、6周时细胞的活性,并应用3.0T磁共振扫描仪对不同浓度标记的细胞行T1、T2序列扫描。结果:MIEG标记EPC后细胞标记率高达98%以上,标记6周后绿色荧光强度有一定程度降低,但荧光标记率仍达90%以上;普鲁士蓝染色显示铁颗粒位于细胞质内。台盼蓝染色结果显示,20 μg/mL MIEG标记EPC对其细胞活性无明显影响(均P>0.05)。体外磁共振扫描结果显示,不同浓度MIEG标记的EPC T1信号强度无明显差异,T2信号强度随MIEG浓度增加而降低。结论:20 μg/mL MIEG标记EPC为适宜的标记浓度,不仅能够高效标记EPC,而且不会影响EPC的细胞活性。磁共振成像体外扫描T2序列可敏感地发现不同浓度MIEG标记EPC后信号强度变化。

关键词: 磁共振成像内皮血管/细胞学同位素标记培养的生物学标记干细胞/细胞学细胞骨髓细胞/细胞学铁化合物/代谢    
Abstract:

Objective: To investigate the feasibility of labeling endothelial progenitor cells (EPCs) with a novel dual modal contrast agent Molday IONTM EverGreen(MIEG) and its performance in vitro MRI.Methods: EPCs were isolated from rat bone marrow and labeled with 10, 20, 50 μg/mL MIEG, respectively. The labeling rates were identified by Prussian blue staining and fluorescence microscopy. The vitality of EPCs labeled with 20 μg/mL MIEG was detected by trypan blue exclusion test at 1 d, 1 w, 2 w, and 6 w after labeling. EPCs labeled with different concentrations of MIEG were scanned by 3.0T MRI with T1 weighted and T2 weighted imaging.Results: The labeling rates for EPCs labeled with different concentrations of MIEG were greater than 98%,and the cytoplasm of labeled EPCs was present with Prussian blue staining. Although the green lighting level went down, the labeling rate at 6 w after labeling was greater than 90%. Trypan blue exclusion test showed that there was no significant difference in the vitality between EPCs labeled with MIEG at 1 d, 1 w, 2 w and 6 w after labeling and EPCs without labeling (all P>0.05). There was no difference in signal intensity on T1 weighted image among EPCs labeled with different concentrations of MIEG. However, the signal intensity on T2 weighted image was reduced in all labeled groups, and the signal reduction became more apparent with increased concentration of MIEG. Conclusions: EPCs can be effectively labeled by MIEG without interference on the cell viability at the labeled concentration of 20 μg/mL. The signal intensity change of labeled cells can be detected sensitively by T2 weighted imaging at 3.0T MRI.

Key words: Magnetic resonance imaging    Endothelium,vascular/cytology    Iron compounds/metabolism    Cells,cultured    Stem cells/cytology    Biological markers    Bone marrow cells/cytology    Isotope labeling
收稿日期: 2017-02-06 出版日期: 2017-10-25
CLC:  R445.2  
基金资助:

国家自然科学基金(30670603);浙江省医药卫生科技计划(2014PYA009)

通讯作者: 陈峰(1961-),男,博士,主任医师,博士生导师,主要从事医学分子影像学研究;E-mail:chenfenghz@zju.edu.cn;http://orcid.org/0000-0003-4402-4955     E-mail: chenfenghz@zju.edu.cn
作者简介: 杨荣(1985-),女,硕士研究生,主治医师,主要从事医学影像诊断工作及医学分子影像学研究;E-mail:dryangrong@zju.edu.cn;http://orcid.org/0000-0002-2322-6200
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引用本文:

杨荣 等. Molday IONTM EverGreen标记大鼠骨髓内皮祖细胞及体外磁共振成像研究[J]. 浙江大学学报(医学版), 2017, 46(5): 481-486.

YANG Rong, YAO Linpeng, ZHANG Siying, CHEN Feng. Rat endothelial progenitor cells labeled with Molday IONTM EverGreen and in vitro MRI study. Journal of ZheJiang University(Medical Science), 2017, 46(5): 481-486.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2017.10.05        http://www.zjujournals.com/xueshu/med/CN/Y2017/V46/I5/481

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