PEI2k-SPIO纳米颗粒标记大鼠骨髓间充质干细胞的体外磁共振成像  

MR Imaging of Polyethylenimine-superparamagnetic Iron Oxide Nanoparticle Labeled Bone Marrow Mesenchymal Stem Cells in vitro

作　　者：陈荟竹[1] 郭应坤[1] 李真林[2] 夏睿[2] 张丽芝[2] 侯江龙[3] 郜发宝[2] 艾华[4] 宁刚[1] 

机构地区：[1]四川大学华西第二医院放射科,成都610041 [2]四川大学华西医院放射科,成都610041 [3]四川大学华西医院胸心外科,成都610041 [4]四川大学生物材料中心,成都610064

出　　处：《四川大学学报（医学版）》2012年第4期578-583,共6页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81130027);四川省科技支撑项目(No.2010SZ0173;2011SZ0108);四川省卫生厅项目(No.100381);成都市科技局项目(No.11DXYB111SF)资助

摘　　要：目的明确经修饰的多聚乙烯-超顺磁性氧化铁(PEI2k-SPIO)纳米颗粒体外标记大鼠骨髓间充质干细胞(BMSCs)的最佳浓度,以及标记后BMSCs的磁共振成像(MRI)特征、可成像的最低标记细胞量和最佳成像细胞量。方法取第二代大鼠BMSCs接种于投放有盖玻片的6孔板内,分别加入浓度为5μg/mL、7μg/mL、10μg/mL、15μg/mL、20μg/mL铁浓度的PEI2k-SPIO液,而后行普鲁士兰染色,观察不同标记浓度对细胞生物学活性的影响;采用四唑盐(MTT)比色法,分析不同浓度的PEI2k-SPIO液对大鼠BMSCs生长活性的影响,并确定PEI2k-SPIO纳米复合颗粒标记干细胞的最佳阈值;取最佳阈值的PEI2k-SPIO标记BMCSs,经磁共振成像,确定成像所需最低细胞量及最佳成像细胞量。结果 PEI2k-SPIO标记BMSCs的最佳浓度是7μg/mL,该标记浓度下细胞生物活性不受影响;在7μg/mL PEI2k-SPIO培养液标记浓度下,MRI的最低细胞量为1×104,最佳成像细胞量为1×106。结论适当浓度的PEI2k-SPIO标记BMSCs对其生物学活性没有影响,MRI可敏感显像PEI2k-SPIO标记的BMSCs。Objective To explore the optimal concentration of polyethylenimine-superparamagnetic iron oxide（PEI2k-SPIO） particles for labeling bone marrow mesenchymal stem cells（BMSCs） in vitro,then to demonstrate the imaging characteristics of the cells by 7.0-T MR scanner.The lowest cell quantity and the optimal cell quantity detected on MR was observed.Methods Cells at 2nd passage were inoculated into the 6-hole plate with cover glass.The different concentrations of PEI2k-SPIO（5 μg/mL,7 μg/mL,10 μg/mL,15 μg/mL,20 μg/mL） were added into different holes,respectively.After labeled with different concentrations of PEI2k-SPIO,the Prussian blue stain was used for determining the labeling efficiency.MTT growth curves were used to identify the activity of BMSCs and to determine the optimal threshold of SPIO nanocomposite particles labeled the stem cells at different PEI2k-SPIO concentrations（7,10,15,20 μg Fe/mL medium）.To definite the lowest cells quantity and the optimal observable cells quantity on MR imaging,1×10^6,1×10^5,1×10^4 and 1×10^3 cells labeled with optimal threshold of PEI2k-SPIO and 1×10^6 cells unlabeled suspended in 0.2 mL agarose（10 g/L）,respectively undergone MR scan.Results MTT growth curves showed the optimal threshold of PEI2k-SPIO labeled BMSCs was 7 μg/mL,which indicates has no adverse effects on the growth of stem cells.At the opimal concentration（7 μg Fe/mL）,the lowest observable cell quantity of PEI2k-SPIO-labeled cells for MRI was 1×10^4,and the optimal observable cell quantity was 1×10^6.Conclusion At the opimal concentration,adverse effect to stem cell activities had not be detected when were labeled with PEI2k-SPIO and the clearly image of MRI of labeled BMSCs could be obtained.

关 键 词：骨髓间充质干细胞 超顺磁性氧化铁 磁共振成像 

分 类 号：R445.2[医药卫生—影像医学与核医学]