Carboxylated superparamagnetic iron oxide particles label cells intracellularly without transfection agents
Carboxylated superparamagnetic iron oxide particles label cells intracellularly without transfection agents
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DOI：
10.1007/s11307-007-0130-3
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发表时间：
2008-05-01
影响因子：
3.1
通讯作者：
Schrezenmeier, Hubert
中科院分区：
文献类型：
--
作者：
Mailaender, Volker;Lorenz, Myriam Ricarda;Schrezenmeier, Hubert
超顺磁性氧化铁颗粒（SPIO）的细胞标记已成为磁共振断层扫描监测移植细胞运输的潜在强大工具，例如在组织修复研究中。然而，细胞内标记主要是通过未被批准用于临床的转染剂来实现的。在这项工作中，评估了两种市售spio (Resovist （R)和Feridex (R)）在不使用转染剂的情况下标记人间充质干细胞（MSC）和HeLa细胞的可行性和效率。在两种细胞类型中，不含转染剂的Resovist (R)比Feridex (R)更有效地被吸收。增加Resovist (R)的浓度可以在细胞中产生与带有转染剂的spio相似的铁量。当使用Resovist (R)时，这提供了从标记方案中省略转染剂的机会。造影剂的细胞内定位是通过光镜发现的，并通过电子显微镜证实。SPIO纳米颗粒的凝固，这是定量细胞内铁含量的问题，用荧光活化细胞分选仪观察和分析。由于Resovist (R)与Feridex (R)是由一个葡聚糖壳构成的，因此我们采用微乳液法合成了具有不同羧基数量的荧光聚合物纳米粒子作为模型体系。随着羧基密度的增加，纳米颗粒的吸收也在稳步增加，这表明在Resovist (R)的情况下，带电基团的相关性。没有发现这些聚合纳米颗粒聚集。
Cell labeling by superparamagnetic iron oxide particles (SPIO) has emerged as a potentially powerful tool to monitor trafficking of transplanted cells by magnetic resonance tomography, e.g., in studies for tissue repair. However, intracellular labeling is mostly achieved by transfection agents not approved for clinical use. In this work, the feasibility and efficiency of labeling human mesenchymal stem cells (MSC) and HeLa cells with two commercially available SPIOs (Resovist (R) and Feridex (R)) without transfection agents was evaluated. In both cell types, Resovist (R) without a transfection agent was more efficiently taken up than Feridex (R). Increasing the concentration of Resovist (R) can yield similar amounts of iron in cells as SPIOs with transfection agents. This offers the opportunity to omit transfection agents from the labeling protocol when Resovist (R) is used. Intracellular localization of the contrast agents is found by light microscopy and confirmed by electron microscopy. Coagulation of the SPIO nanoparticles, which is problematic for the quantification of the intracellular iron content, was observed and analyzed with a fluorescent activated cell sorter. As Resovist (R) consists of a carboxydextran shell in contrast to Feridex (R) which is composed of a dextran shell, we synthesized fluorescent polymeric nanoparticles as model systems with different amounts of carboxyl groups on the surface by the miniemulsion process. A steady increase in uptake of nanoparticles was detected with a higher density of carboxyl groups showing the relevance of charged groups as in the case of Resovist (R). Aggregation of these polymeric nanoparticles was not found.