| 郑敏文,宦怡,徐健,葛雅丽,赵力,程康,尹涛,孙立军,赵海涛.超顺磁性氧化铁标记骨髓间充质干细胞的磁共振成像研究[J].中国医学影像技术,2006,22(8):1129~1134 |
| 超顺磁性氧化铁标记骨髓间充质干细胞的磁共振成像研究 |
| In vitro MR imaging of superparamagnetic iron oxide labeled mesenchymal stem cells |
| 投稿时间：2006-04-12 修订日期：2006-05-30 |
| DOI： |
| 中文关键词: 干细胞 移植 超顺磁性氧化铁 磁共振成像 |
| 英文关键词:Stem cells Transplantation Superparamagnetic iron oxid Magnetic resonance imaging |
| 基金项目: |
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| 中文摘要: |
| 目的 明确顺磁性氧化铁纳米粒子(SPIO)体外标记兔骨髓间充质干细胞(MSCs)的适当浓度和不同标记浓度对细胞的生物学活性影响,以及经MR成像的特征和可成像的最低标记细胞量等。方法 分离、纯化、培养兔MSCs,体外不同浓度SPIO标记,荧光显微镜观察铁颗粒在细胞内的位置、不同标记浓度下的最佳孵育时间、不同标记浓度细胞形态的改变、细胞内铁颗粒的分布及标记率;测量并绘制未标记细胞和标记细胞的MTT生长曲线;选取适当浓度标记量,对不同细胞量组进行标记后MR成像,测量不同扫描序列标记细胞管的信号强度改变,并进行统计学分析。结果 标记后的铁颗粒均位于细胞质内;20~50 μg Fe/ml培养液是SPIO标记干细胞的适当浓度阈值,超过50 μg Fe/ml培养液浓度可使细胞的变形增殖能力受到不同程度的抑制;在此浓度阈值标记后孵育18~24 h即可有效标记细胞97%~100％;细胞对铁颗粒的吸收与细胞的数量、标记浓度和孵育时间呈正相关;SPIO标记的MSCs在T2WI尤其是FFE(T2*WI)序列信号明显降低;在35 μg Fe/ml培养液标记浓度下,MR可成像的最低细胞量为5×104;35 μg Fe/ml培养液浓度时,标记细胞1×105和5×104 MR成像可使T2WI、FFE图像信号均降低,而且没有使靶灶扩大的磁敏感伪影。结论 SPIO可以简便标记MSCs并且在适当浓度下对MSCs的生物学活性没有影响,MR T2WI和T2*WI序列可敏感显像磁性标记的干细胞。 |
| 英文摘要: |
| Objective To in vitro evaluate the labeling efficiency of rabbit mesenchymal stem cells (MSCs) with different labeling concentrations of superparamagnetic iron oxide (SPIO) nanoparticles, and to detect the characteristics and signal attenuation rules by a 1.5T MR scanner. Methods MSCs were isolated from rabbit and incubated with different concentrations of SPIO particles at 37℃ in 5% CO2. To measure the labeling efficiency of SPIO, the samples labeled at a range of SPIO concentrations separately were observed and counted under fluorescent microscope for evidence of distribution and labeling ratio of SPIO particles in cells, optimal incubation time after labeling and morphological evidence of abnormal visualization. To assess the effects of the particles on cell proliferation, MTT growth curves were obtained at a range of SPIO concentrations (18 to 210 μg Fe per ml medium). At an optimal concentration, samples of SPIO-labeled and unlabeled MSCs suspension in agar were imaged by MRI with T1WI, T2WI and fast field echo (FFE) sequences, and the signal intensity were measured and statistically analyzed. Results Labeled SPIO particles were stained in cytoplasm. MSCs were efficiently labeled (97% to 100% by fluorescent microscope) in culture at a range of exposure time (from 18 to 24 hours) and particle concentrations (20 to 50 μg Fe per ml medium). Compared to unlabeled cells, MSCs loaded with SPIO had similar viability and proliferation profiles at this proper range of labeling concentration. Endocytosis of iron particles were strongly depended on the cell quantity of MSCs in media, cellular labeling concentration, and incubation time. SPIO labeling caused a stronger low signal attenuation effect in FFE and T2WI than in T1WI. At a labeling concentration of 35 μg Fe per ml medium, MRI demonstrated that 105 SPIO-labeled cells/ml medium was the lowest observable cells quantity. 5×104 to 1×105 SPIO-labeled cells/ml medium were able to decrease the signal without magnetic sensitivity artificials in both T2WI and FFE. Conclusion MSCs can be easily and efficiently labeled by SPIO without interference on the cell viability and proliferation, MRI visualization of SPIO-labeled MSCs is feasible, which may be critical for future experimental studies. |
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