[1]杨华,张小明,邵阳,等.SPIO标记大鼠骨髓基质干细胞及体外磁共振成像研究[J].陆军军医大学学报(原第三军医大学学报),2008,30(17):1626-1629.
　YANG Hua,ZHANG Xiao-ming,SHAO Yang,et al.Superparamagnetic iron oxide labeled mesenchymal stem cells and their magnetic resonance imaging in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(17):1626-1629.
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第17期
页码:
1626-1629
栏目:
论著
出版日期:
2008-09-15

文章信息/Info

Title:
Superparamagnetic iron oxide labeled mesenchymal stem cells and their magnetic resonance imaging in vitro
作者:
杨华; 张小明; 邵阳; 蒋红; 翟昭华
第三军医大学新桥医院放射科；川北医学院附属医院: 放射科， 风湿免疫研究所
Author(s):
YANG Hua;  ZHANG Xiao-ming;  SHAO Yang;  JIANG Hong;  ZHAI Zhao-hua
Department of Radiology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037;Department of Radiology, 3Department of Immunology, The Affiliated Hospital of North Sichuan Medical College, Nanchong Sichuan 637000, China
关键词:
超顺磁性氧化铁; 多聚赖氨酸; 骨髓基质干细胞; 磁共振成像
Keywords:
superparamagnetic iron oxides;  poly-L-lysine;  bone mesenchymal stem cells;  magnetic resonance image
分类号:
R331.22; R445.2; R817.1
文献标志码:
A
摘要:
目的    探讨以多聚赖氨酸(poly-L-lysine，PLL)为转染介质介导超顺磁性氧化铁微粒(superparamgnetic iron oxides，SPIO)标记大鼠骨髓基质干细胞(bone mesenchymal stem cells，BMSCs)的合适条件,通过标记细胞的体外磁共振成像（magnetic resonance imaging,MRI）获取最佳的磁共振扫描序列。    方法    分离、纯化、培养大鼠BMSCs ,SPIO和PLL以1∶0.001 2 混合配制氧化铁-多聚赖氨酸(F-PLL)混合液，用铁浓度分别为0、4.2、8.4、21、42、84 μg/ml的FE-PLL复合物标记BMSCs。计算细胞标记阳性率，测量并绘制未标记细胞和不同浓度铁标记细胞的MTT生长曲线;对不同浓度铁标记的细胞进行磁共振成像。分别用浓度为0、0.05、0.25、0.5、1.0、5.0 μg/ml的PLL同BMSCs共同培养，了解各种浓度PLL对细胞的生存是否有影响。    结果    细胞的标记率分别为0％、（44.40±11.33）％、（71.97±8.01）％、（88.86±9.10）％、（98.24±2.94）％、100％，随着SPIO浓度的增加而增加。MTT显示在铁浓度<42 μg/ml时FE-PLL复合物对细胞的生长活性无明显影响，而铁浓度为84 μg/ml时，细胞的生长活性受到抑制。PLL在浓度≤1.0 μg/ml时对细胞的活力无明显影响；当PLL浓度为5.0 μg/ml时，细胞生长明显受抑。MRI信号随FE-PLL复合物浓度的增加而增加(P<0.05)，标记细胞的信号在T1WI的改变不及在T2WI及T2*WI明显，而以T2*WI信号改变最明显(P<0.05)。    结论    以PLL为转染介质，SPIO能够高效的标记BMSCs，并且在合适的浓度范围内不会影响干细胞的生长活性。
Abstract:
Objective    To explore the optimal situation of labeling bone mesenchymal stem cells (BMSCs) with superparamagnetic iron oxides (SPIO) mediated by poly-L-lysine (PLL), and determine the most optimal protocol of magnetic resonance imaging according to the patterns of MR in vitro.     Methods    BMSCs were isolated from white rat and purified, incubated with SPIO-PLL complexes at the range of concentrations (0, 4.2, 8.4, 21, 42, 84 μg Fe per ml medium). The labeling ratio and distribution of SPIO particles in BMSCs, and the morphological evidence of abnormal visualization were evaluated by Prussian blue staining, fluorescent microscope and electron microscopy. MTT growth curves and magnetic resonance imagings were obtained at the range of concentrations. Trypan blue exclusion test was performed to elevate the viability of BMSCs labeled with PLL at the range of concentrations (0, 0.05, 0.25, 0.5, 1.0, 5.0 μg PLL per ml medium).     Results    The cellular labeling ratio was strongly correlated to the concentrations of SPIO (P<0.05). BMSCs labeled with SPIO had similar viability and proliferation profiles at the range of 0 to 42 μg Fe per ml medium, while the cellular viability was limited at the concentration of 84 μg/ml (P<0.05). PLL could not affect the cellular viabilities when its concentration was in the range of 0 to 1.0 μg PLL per ml medium, but the viabilities of BMSCs were significantly limited at the concentration of 5.0 μg/ml of PLL. The signal intensities of cell suspension were strongly dependent on the concentrations of SPIO used for labeling in medium (P<0.05). The signal intensity rates on T2WI and T2*WI were stronger than that on T1WI (P<0.05), especially on T2*WI.      Conclusion    SPIO can label BMSCs easily and efficiently mediated by PLL without interference on the cellular viability and proliferation. The optimal concentration of SPIO should range from 20 μg/ml to 40 μg/ml when incubation time was 24 h.

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更新日期/Last Update: 2008-09-16