Pawelczyk Edyta, Arbab Ali S, Pandit Sunil, Hu Elbert, Frank Joseph A
Laboratory of Diagnostic Radiology Research, Experimental Neuroimaging Section, Clinical Center, National Institutes of Health, Bethesda, MD 20892, USA.
NMR Biomed. 2006 Aug;19(5):581-92. doi: 10.1002/nbm.1038.
Ferumoxides-protamine sulfate (FE-Pro) complexes are used for intracellular magnetic labeling of cells to non-invasively monitor cell trafficking by in vivo MRI. FE-Pro labeling is non-toxic to cells; however, the effects of FE-Pro labeling on cellular expression of transferrin receptor (TfR-1) and ferritin, proteins involved in iron transport and storage, has not been reported. FE-Pro-labeled human mesenchymal stem cells (MSCs), HeLa cells and primary macrophages were cultured from 1 week to 2 months and evaluated for TfR-1 and ferritin gene expression by RT-PCR and protein levels were determined using Western blots. MTT (proliferation assay) and reactive oxygen species (ROS) analysis were performed. FE-Pro labeling of HeLa and MSCs resulted in a transient decrease in TfR-1 mRNA and protein levels. In contrast, Fe-Pro labeling of primary macrophages resulted in an increase in TfR-1 mRNA but not in TfR-1 protein levels. Ferritin mRNA and protein levels increased transiently in labeled HeLa and macrophages but were sustained in MSCs. No changes in MTT and ROS analysis were noted. In conclusion, FE-Pro labeling elicited physiological changes of iron metabolism or storage, validating the safety of this procedure for cellular tracking by MRI.
硫酸铁氧化物-鱼精蛋白(FE-Pro)复合物用于细胞的细胞内磁性标记,以通过体内磁共振成像(MRI)非侵入性地监测细胞运输。FE-Pro标记对细胞无毒;然而,FE-Pro标记对转铁蛋白受体(TfR-1)和铁蛋白(参与铁运输和储存的蛋白质)细胞表达的影响尚未见报道。对FE-Pro标记的人间充质干细胞(MSCs)、HeLa细胞和原代巨噬细胞进行1周至2个月的培养,并通过逆转录聚合酶链反应(RT-PCR)评估TfR-1和铁蛋白基因表达,使用蛋白质印迹法测定蛋白质水平。进行了MTT(增殖测定)和活性氧(ROS)分析。HeLa细胞和MSCs的FE-Pro标记导致TfR-1 mRNA和蛋白质水平短暂下降。相反,原代巨噬细胞的Fe-Pro标记导致TfR-1 mRNA增加,但TfR-1蛋白质水平未增加。铁蛋白mRNA和蛋白质水平在标记的HeLa细胞和巨噬细胞中短暂增加,但在MSCs中持续增加。MTT和ROS分析未见变化。总之,FE-Pro标记引发了铁代谢或储存的生理变化,验证了该程序用于MRI细胞追踪的安全性。