SPIO标记大鼠骨髓间充质干细胞:生物学活性及体外MRI成像效应评价  

SPIO-labeled Rat Bone Marrow Mesenchymal Stem Cells:Alterations of Biological Activity and Labeling Efficiency Assay In Vitro

作　　者：吕鑫[1] 聂永梅[1] 赵志伟[2] 何学令[3] 刘艳[3] 甫拉提.吐尔逊 吴江[1,2] 

机构地区：[1]新疆医科大学基础医学院,乌鲁木齐830011 [2]四川大学华西基础与法医学院,成都610041 [3]四川大学实验动物中心,成都610041

出　　处：《生物医学工程学杂志》2014年第2期365-372,共8页Journal of Biomedical Engineering

基　　金：国家自然科学基金项目资助课题(11262020);新疆维吾尔自治区自然科学基金资助项目(201233146-9);四川省应用基础计划项目(2013JY0021)

摘　　要：为研究不同浓度（25、50、100 μg/mL）超顺磁性氧化铁粒子（SPIO）标记对大鼠骨髓间充质干细胞（rMSCs）生物学活性、体外MRI成像效应的影响，采用多聚赖氨酸（PLL）包被的SPIO（PLL-SPIO）标记原代分离培养的rMSCs，24 h后分别进行阳性标记率、MRI成像效应、细胞增殖凋亡以及干细胞细胞分化能力的综合评价。PLL-SPIO标记阳性率、铁含量测定结果显示：铁标记率可达75%~100%。透射电镜观察，可见100 μg/mL组rMSCs细胞质内浓聚细小铁颗粒的囊泡样包涵体。采用T1WI、T2WI和T2＊WI序列MRI成像扫描显示：25 μg/mL组即可引起信号的明显降低。与未标记的对照组细胞相比，25 μg/mL 和50 μg/mL组细胞增殖活力、凋亡、干细胞分化能力检测结果差异皆无统计学意义（P＞0.05），而100 μg/mL组的差异均有统计学意义（P＜0.05）。结论： 25、50 μg/mL SPIO可有效标记rMSCs，100 μg/mL的SPIO对rMSCs增殖活性和分化能力有抑制作用，这为SPIO进一步的体内及临床MRI分子影像应用提供了基础。This study aimed to characterize and magnetic resonance imaging （MRI） track the mesenchymal stem cells labeled with polylysine-coated superparamagnetic iron oxide （PLL-SPIO）. Rat bone marrow derived mesenchymal stem cells （rMSCs） were labeled with 25, 50 and 100 μg/mL PLL-SPIO for 24 hours. The labeling efficiency was assessed by iron content, Prussian blue staining, electron microscopy and in vitro MR imaging. The labeled cells were also analyzed for cytotoxicity and differentiation potential. Electron microscopic observations and Prussian blue staining revealed that 75%-100% of cells were labeled with iron particles. PLL-SPIO did not show any cytotoxicity up to 100 μg/mL concentration. Both 25 μg/mL and 50/zg/mL PLL-SPIO labeled stem cells did not exhibit any sig- nificant alterations in the adipo/osteo/chondrogenic differentiation potential compared to unlabeled control cells. The lower concentration of 25 μg/mL iron labeled cells emitted an obvious dark signal in T1W, T2WI and T2 ＊ WI MR image. The novel PLL-SPIO enables to label and track rMSCs for in vitro MRI without cellular alteration. Therefore PLL-SPIO may potentially become a better MR contrast agent especially in tracking the transplanted stem cells and other cells without compromising cell functional quality.

关 键 词：超顺磁性氧化铁粒子 磁共振成像 骨髓间充质干细胞 

分 类 号：R318.5[医药卫生—生物医学工程] R445.2[医药卫生—基础医学]