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无转染剂情况下间充质干细胞的磁性纳米颗粒标记:单细胞水平的细胞行为及临床1.5T磁共振检测能力

Magnetic nanoparticle labeling of mesenchymal stem cells without transfection agent: cellular behavior and capability of detection with clinical 1.5 T magnetic resonance at the single cell level.

作者信息

Hsiao Jong-Kai, Tai Ming-Fong, Chu Hung-Hao, Chen Shin-Tai, Li Hung, Lai Dar-Ming, Hsieh Sung-Tsang, Wang Jaw-Lin, Liu Hon-Man

机构信息

Department of Medical Imaging, National Taiwan University Hospital, Taipei, Taiwan.

出版信息

Magn Reson Med. 2007 Oct;58(4):717-24. doi: 10.1002/mrm.21377.

Abstract

The purpose of this work was to evaluate the efficacy of labeling human mesenchymal stem cells (hMSCs) by ionic superparamagnetic iron oxide (SPIO) without a transfection agent and verifying its capability to be detected with clinical 1.5 T magnetic resonance (MR) at the single-cell level. Human hMSCs were incubated for 24 h with an ionic SPIO, Ferucarbotran. The labeling efficiency of hMSCs was determined by iron content measurement spectrophotometrically, and the influence of labeling on cell behavior was ascertained by examination of cell viability using the trypan blue exclusion method, cell proliferation analysis using MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, mitochondrial membrane potential (MMP) change, differentiation capacity, and reactive oxygen species (ROS) production measured by dichlorofluorescein diacetate (DCFDA) fluorescent probe. Labeled hMSCs were scanned under 1.5 T MRI with three-dimensional (3D) and two-dimensional (2D) T(2)-weighted gradient echo (GRE) pulse sequences. Human hMSC labeling without transfection agent was efficient. The iron content in hMSCs was 23.4 pg Fe/cell. No significant change was found in viability, proliferation, MMP change, ROS production, or differentiation capacity. About 45.2% of the hMSCs could be detected using 1.5 T MRI at the single cell level with 3D GRE and four repetitions.

摘要

本研究旨在评估在不使用转染剂的情况下,利用离子超顺磁性氧化铁(SPIO)标记人间充质干细胞(hMSCs)的效果,并验证其在临床1.5 T磁共振(MR)下在单细胞水平被检测到的能力。将人hMSCs与离子SPIO Ferucarbotran孵育24小时。通过分光光度法测量铁含量来确定hMSCs的标记效率,并通过台盼蓝排斥法检测细胞活力、使用MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)法进行细胞增殖分析、测量线粒体膜电位(MMP)变化、分化能力以及使用二氯荧光素二乙酸酯(DCFDA)荧光探针测量活性氧(ROS)生成,来确定标记对细胞行为的影响。使用三维(3D)和二维(2D)T(2)加权梯度回波(GRE)脉冲序列在1.5 T MRI下对标记的hMSCs进行扫描。不使用转染剂的人hMSC标记是有效的。hMSCs中的铁含量为23.4 pg Fe/细胞。在活力、增殖、MMP变化、ROS生成或分化能力方面未发现显著变化。使用3D GRE和四次重复扫描,在单细胞水平利用1.5 T MRI可检测到约45.2%的hMSCs。

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