Arbab Ali S, Bashaw Lindsey A, Miller Bradley R, Jordan Elaine K, Lewis Bobbi K, Kalish Heather, Frank Joseph A
Experimental Neuroimaging Section, Laboratory of Diagnostic Radiology Research, National Institutes of Health, 10 Center Dr, Rm B1N256, Bethesda, MD 20892, USA.
Radiology. 2003 Dec;229(3):838-46. doi: 10.1148/radiol.2293021215.
To evaluate the effect of using the ferumoxides-poly-l-lysine (PLL) complex for magnetic cell labeling on the long-term viability, function, metabolism, and iron utilization of mammalian cells.
PLL was incubated with ferumoxides for 60 minutes, incompletely coating the superparamagnetic iron oxide (SPIO) through electrostatic interactions. Cells were coincubated overnight with the ferumoxides-PLL complex, and iron uptake, cell viability, apoptosis indexes, and reactive oxygen species formation were evaluated. The disappearance or the life span of the detectable iron nanoparticles in cells was also evaluated. The iron concentrations in the media also were assessed at different time points. Data were expressed as the mean +/- 1 SD, and one-way analysis of variance and the unpaired Student t test were used to test for significant differences.
Intracytoplasmic nanoparticles were stained with Prussian blue when the ferumoxides-PLL complex had magnetically labeled the human mesenchymal stem and HeLa cells. The long-term viability, growth rate, and apoptotic indexes of the labeled cells were unaffected by the endosomal incorporation of SPIO, as compared with these characteristics of the nonlabeled cells. In nondividing human mesenchymal stem cells, endosomal iron nanoparticles could be detected after 7 weeks; however, in rapidly dividing cells, intracellular iron had disappeared by five to eight divisions. A nonsignificant transient increase in reactive oxygen species production was seen in the human mesenchymal stem and HeLa cell lines. Labeled human mesenchymal stem cells did not differentiate to other lineage. A significant increase in iron concentration was observed in both the human mesenchymal stem and HeLa cell media at day 7.
Magnetic cellular labeling with the ferumoxides-PLL complex had no short- or long-term toxic effects on tumor or stem cells.
评估使用超顺磁性氧化铁-聚-L-赖氨酸(PLL)复合物进行磁性细胞标记对哺乳动物细胞的长期活力、功能、代谢及铁利用情况的影响。
将PLL与超顺磁性氧化铁孵育60分钟,通过静电相互作用不完全包覆超顺磁性氧化铁(SPIO)。细胞与超顺磁性氧化铁-PLL复合物共孵育过夜,评估铁摄取、细胞活力、凋亡指数及活性氧生成情况。还评估了细胞中可检测到的铁纳米颗粒的消失情况或寿命。在不同时间点评估培养基中的铁浓度。数据以平均值±1标准差表示,采用单因素方差分析和非配对学生t检验来检验显著差异。
当超顺磁性氧化铁-PLL复合物对人间充质干细胞和HeLa细胞进行磁性标记后,胞质内纳米颗粒用普鲁士蓝染色。与未标记细胞的这些特征相比,SPIO的内吞作用未影响标记细胞的长期活力、生长速率和凋亡指数。在不分裂的人间充质干细胞中,7周后可检测到内吞的铁纳米颗粒;然而,在快速分裂的细胞中,细胞内铁在五到八次分裂后消失。人间充质干细胞系和HeLa细胞系中活性氧生成有不显著的短暂增加。标记的人间充质干细胞未分化为其他谱系。在第7天,人间充质干细胞和HeLa细胞培养基中的铁浓度均显著增加。
超顺磁性氧化铁-PLL复合物进行磁性细胞标记对肿瘤细胞或干细胞无短期或长期毒性作用。