[用不同的超顺磁性氧化铁颗粒标记间充质干细胞及在3T磁共振成像下的可检测性]

[Labeling of mesenchymal stem cells with different superparamagnetic particles of iron oxide and detectability with MRI at 3T].

作者信息

Ittrich H, Lange C, Dahnke H, Zander A R, Adam G, Nolte-Ernsting C

机构信息

Klinik und Poliklinik für Diagnostische und Interventionelle Radiologie, Radiologisches Zentrum, Universitätsklinikum Hamburg-Eppendorf.

出版信息

Rofo. 2005 Aug;177(8):1151-63. doi: 10.1055/s-2005-858330.

DOI:10.1055/s-2005-858330

PMID:16021549

Abstract

PURPOSE

In vitro evaluation of labeling efficiency of human mesenchymal stem cells (hMSCs) with different types of superparamagnetic iron oxide nanoparticles as well as detection and quantification by MRI at 3T.

MATERIAL AND METHODS

hMSCs were incubated for 24 hours with 5 ultrasmall superparamagnetic particles of iron oxide (USPIO) contrast agents (1 : 30 - 1 : 30,000) of different size, coating and core compound: Endorem, Resovist, citric acid coated magnetite cores of 3 nm (CMF3), 7 nm (CMF7) and 12 nm (CoF, core: cobalt ferrite). Iron uptake, intracellular retention, detection and quantification were evaluated with MRI up to 5 weeks after incubation by cytological analysis (Prussian blue), atomic absorption spectrometry and MR relaxometry measurements.

RESULTS

An effective labeling of hMSCs was achieved using Resovist, CMF3 and CMF7 with mean iron concentrations of 5.1/1.8, 1.9/1.4 and 1.5/1.0 pg/cell (dilutions 1:30 [933, 2100, 2800 microg Fe/ml]/1 : 300 [93, 210, 280 microg Fe/ml]) compared with 0.58/0.34 and 0.43/0.30 pg/cell (Endorem, CoF, dilution 1 : 30 [400, 4200 microg Fe/ml]/1 : 300 [40, 420 microg Fe/ml] unlabelled control cells: 0.01 pg/cell). Particle uptake correlated with the concentration of USPIO in the incubation medium. Detection of 5 x 10 (4) labelled cells/ml with MRI was possible up to 5 weeks after incubation (Resovist, CMF7 and CMF3). MR relaxometry measurements showed a strong correlation between cellular iron load and R2* (1/T2*), r > 0.78. No changes in cell viability or toxic effects were found.

CONCLUSION

Efficiency of labeling hMSCs with USPIOs depends on coating, size and core compound of used particles. Carboxydextran-coated, clinically approved SPIO (Resovist, 50 nm) or ultrasmall citrate-coated particles (< 10 nm) result in an improved cellular uptake. In principle, the long intracellular retention of particles offers the possibility of cell tracking and migration monitoring in MRI.

摘要

目的

在体外评估不同类型的超顺磁性氧化铁纳米颗粒对人间充质干细胞（hMSCs）的标记效率，并通过3T磁共振成像（MRI）进行检测和定量分析。

材料与方法

将hMSCs与5种不同尺寸、包被和核心化合物的超小超顺磁性氧化铁（USPIO）造影剂（1:30 - 1:30,000）孵育24小时：Endorem、Resovist、3nm柠檬酸包被的磁铁矿核心（CMF3）、7nm（CMF7）和12nm（CoF，核心：钴铁氧体）。在孵育后长达5周的时间内，通过细胞学分析（普鲁士蓝）、原子吸收光谱法和磁共振弛豫测量法，利用MRI评估铁摄取、细胞内保留、检测和定量分析。

结果

使用Resovist、CMF3和CMF7可有效标记hMSCs，平均铁浓度分别为5.1/1.8、1.9/1.4和1.5/1.0 pg/细胞（稀释度1:30 [933, 2100, 2800 μg Fe/ml]/1:300 [93, 210, 280 μg Fe/ml]），而Endorem和CoF的平均铁浓度分别为0.58/0.34和0.43/0.30 pg/细胞（稀释度1:30 [400, 4200 μg Fe/ml]/1:300 [40, 420 μg Fe/ml]，未标记对照细胞：0.01 pg/细胞）。颗粒摄取与孵育培养基中USPIO的浓度相关。孵育后长达5周，通过MRI能够检测到5×10⁴个标记细胞/ml（Resovist、CMF7和CMF3）。磁共振弛豫测量显示细胞铁负荷与R2*（1/T2*）之间存在强相关性，r > 0.78。未发现细胞活力变化或毒性作用。