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目的评估超小超顺磁性氧化铁(ultrasmall superparamagnetic iron oxide,USPIO)标记SD大鼠脂肪来源干细胞(adipose derived stem cells,ADSCs)的有效性及安全性,探讨标记细胞体外磁共振成像(magnetic resonance ima-ging,MRI)特点。方法将USPIO40μg/ml及多聚赖氨酸(poly-L-lysine,PLL)1·5μg/ml的培养基与ADSCs共孵育培养24h,检测USPIO标记的有效性及安全性,并用MRI对标记细胞进行体外成像。结果普鲁士蓝染色显示USPIO标记ADSCs的阳性率为99%,透射电镜提示USPIO颗粒主要位于胞质内溶酶体中;台盼蓝染色实验显示活细胞数大于95%,MTS[3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,溴化噻唑蓝四氮唑]实验提示USPIO浓度为10、20、40、80和160μg/ml时不影响ADSCs增殖。ELISA结果表明标记细胞与未标记细胞组间培养液中血管内皮生长因子(vascular endothelial growth factor,VEGF)水平无显著差异;体外条件下,MRI图像信号强度与标记细胞数量呈正相关。结论 USPIO标记ADSCs安全有效,MRI图像信号强度与标记细胞数量存在一定相关性,提示USPIO可用于在体条件下MRI成像示踪标记细胞。
Objective To assess the efficacy and safety of adipose derived stem cells (ADSCs) labeled with ultrasmall superparamagnetic iron oxide (USPIO) in SD rats and to investigate the effect of magnetic resonance imaging -ging, MRI) features. Methods The culture media of 40 μg / ml USPIO and 1.5 μg / ml poly-L-lysine (PLL) were co-incubated with ADSCs for 24 h. The efficiency and safety of USPIO labeling were detected. In vitro imaging. Results Prussian blue staining showed that the positive rate of USPIO-labeled ADSCs was 99%. Transmission electron microscopy suggested that USPIO particles were mainly localized in the cytoplasmic lysosomes. Trypan blue staining showed that the number of viable cells was more than 95%. MTS [3- (4, 5-dimethylthiazol-2-yl) -5 (3-carboxymethoxyphenyl) -2- (4-sulfopheny) -2H-tetrazolium showed that USPIO concentrations were 10, 20, 40, 80 and 160 μg / ml does not affect ADSCs proliferation. The results of ELISA showed that there was no significant difference in the level of vascular endothelial growth factor (VEGF) between the labeled and unlabeled cells. In vitro, the signal intensity of MRI was positively correlated with the number of labeled cells. Conclusions The USPIO labeled ADSCs is safe and effective. The signal intensity of MRI is correlated with the number of labeled cells, suggesting that USPIO can be used to label the labeled cells under MRI in vivo.