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聚乙烯亚胺-超顺磁性氧化铁纳米粒子标记的骨髓间充质干细胞的体外磁共振成像

[MR imaging of polyethylenimine-superparamagnetic iron oxide nanoparticle labeled bone marrow mesenchymal stem cells in vitro].

作者信息

Chen Hui-Zhu, Guo Ying-Kun, Li Zhen-Lin, Xia Rui, Zhang Li-Zhi, Hou Jiang-Long, Gao Fa-Bao, Ai Hua, Ning Gang

机构信息

Department of Radiology, West China Second Hospital, Sichuan University, Chengdu 610041, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2012 Jul;43(4):578-83.

Abstract

OBJECTIVE

To explore the optimal concentration of polyethylenimine-superparamagnetic iron oxide (PEI2k-SPIO) particles for labeling bone marrow mesenchymal stem cells (BMSCs) in vitro, then to demonstrate the imaging characteristics of the cells by 7.0-T MR scanner. The lowest cell quantity and the optimal cell quantity detected on MR was observed.

METHODS

Cells at 2nd passage were inoculated into the 6-hole plate with cover glass. The different concentrations of PEI2k-SPIO (5 microg/mL, 7 microg/mL, 10 microg/mL, 15 microg/mL, 20 microg/mL) were added into different holes, respectively. After labeled with different concentrations of PEI2k-SPIO, the Prussian blue stain was used for determining the labeling efficiency. MTT growth curves were used to identify the activity of BMSCs and to determine the optimal threshold of SPIO nanocomposite particles labeled the stem cells at different PEI2k-SPIO concentrations (7, 10, 15, 20 microg Fe/mL medium). To definite the lowest cells quantity and the optimal observable cells quantity on MR imaging, 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells labeled with optimal threshold of PEI2k-SPIO and 1 x 10(6) cells unlabeled suspended in 0.2 mL agarose (10 g/L), respectively undergone MR scan.

RESULTS

MTT growth curves showed the optimal threshold of PEI2k-SPIO labeled BMSCs was 7 microg/mL, which indicates has no adverse effects on the growth of stem cells. At the opimal concentration (7 microg Fe/mL), the lowest observable cell quantity of PEI2k-SPIO-labeled cells for MRI was 1 x 10(4), and the optimal observable cell quantity was 1 x 10(6).

CONCLUSION

At the opimal concentration, adverse effect to stem cell activities had not be detected when were labeled with PEI2k-SPIO and the clearly image of MRI of labeled BMSCs could be obtained.

摘要

目的

探讨聚乙烯亚胺-超顺磁性氧化铁(PEI2k-SPIO)颗粒体外标记骨髓间充质干细胞(BMSCs)的最佳浓度,然后用7.0-T MR扫描仪显示细胞的成像特征。观察MR检测到的最低细胞数量和最佳细胞数量。

方法

将第2代细胞接种到带有盖玻片的6孔板中。分别向不同孔中加入不同浓度的PEI2k-SPIO(5μg/mL、7μg/mL、10μg/mL、15μg/mL、20μg/mL)。用不同浓度的PEI2k-SPIO标记后,采用普鲁士蓝染色法测定标记效率。MTT生长曲线用于鉴定BMSCs的活性,并确定不同PEI2k-SPIO浓度(7、10、15、20μg Fe/mL培养基)下SPIO纳米复合颗粒标记干细胞的最佳阈值。为确定MR成像上的最低细胞数量和最佳可观察细胞数量,分别将1×10⁶、1×10⁵、1×10⁴和1×10³个用PEI2k-SPIO最佳阈值标记的细胞以及1×10⁶个未标记的细胞悬浮于0.2 mL琼脂糖(10 g/L)中进行MR扫描。

结果

MTT生长曲线显示PEI2k-SPIO标记BMSCs的最佳阈值为7μg/mL,这表明对干细胞生长无不良影响。在最佳浓度(7μg Fe/mL)下,PEI2k-SPIO标记细胞用于MRI的最低可观察细胞数量为1×10⁴,最佳可观察细胞数量为1×10⁶。

结论

在最佳浓度下,用PEI2k-SPIO标记时未检测到对干细胞活性的不良影响,并且可以获得标记BMSCs的清晰MR图像。

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