体外磁铁定向迁移SPIO标记的间充质干细胞对大鼠脑缺血治疗的影响

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (10): 1302-1308.

体外磁铁定向迁移SPIO标记的间充质干细胞对大鼠脑缺血治疗的影响

郑洁,陈晓丹,郑希,王顺和

重庆医科大学

收稿日期:2012-10-17 修回日期:2013-03-26 出版日期:2013-10-05 发布日期:2013-09-25
通讯作者: 王顺和 E-mail:wangshunhe120@yahoo.com

The effects of migration of superparamagnetic iron oxide (SPIO)-Labeled mesenchymal stem cells by a magnet on brain infarction in rat model

Received:2012-10-17 Revised:2013-03-26 Online:2013-10-05 Published:2013-09-25

摘要/Abstract

摘要： 目的探讨外加磁场定向迁移SPIO标记的大鼠骨髓间充质干细胞(BMSCs)对移植治疗大鼠脑缺血的影响。方法体外培养BMSCs，SPIO标记BMSCs，用普鲁士蓝染色测定标记率；台盼蓝染色测定细胞存活率；CCK法检测磁场对标记干细胞的增殖活力。建立大鼠脑缺血模型，用尾静脉注射移植细胞；对移植后大鼠进行神经学评估；普鲁士蓝染色观察脑组织内标记细胞；TTC染色观察脑缺血面积；分光光度计检测脑组织铁含量。结果细胞标记率接近100%，普鲁士蓝染色观察磁性标记BMSCs静脉移植并加外加磁场组（ICSM组）脑组织内BMSCs明显多于未磁性标记BMSCs静脉移植组（ICS组）（P<0.05），移植后7d后ICSM组动物神经系统行为学评分明显改善(P<0.05)，ICSM组脑组织内铁含量明显多于ICS组（P<0.05）。结论在体内和体外使用外加磁铁均可以定向迁移标记的BMSCs，明显的提高经尾静脉移植标记SPIO的BMSCs的成功率。

关键词: 关键词：磁场, 骨髓间充质干细胞, 超顺次氧化铁, 脑缺血, 迁移

Abstract: Objective To observe the effect of rat bone marrow mesenchymal stem cells(BMSCs) labeled with superparamagnetic iron oxide(SPIO) in the magnetic field on the transplantation for rat focal cerebral ischemia. Methods The rat mesenchymal stem cells was labeled with SPIO in vitro , and prussian blue staining was used to check the rate of SPIO. The viability and proliferation activity of labeled stem cells was evaluated by trypan blue staining. The cck-8 assay was performed to detect the labeled stem cell proliferation activity in the magnetic field. The middle cerebral artery occlusion (MCAO) was established in a rat model, and the neurological severity score (NSS) was used to evaluate neurological function recovery of rats at definite time following the injection of transplanted cells. The transplanted BMSCs were analyzed in recipient rat brains by Prussian blue staining. The cerebral ischemia area was observed via TTC staining. The iron content of SPIO-labeled BMSCs was analyzed by atomic absorption spectrophotometer. Results Prussian blue staining showed that the rate of BMSCs labeled superparamagnetic iron oxide was close to 100％. Trypan blue staining indicated that there was no significant difference between labeled group and control group （P>0.05）. CCK-8 assay suggest that there was no significant difference between magnetic group and control group （P>0.05）. The BMSCs of ICSM group within the brain was significantly more than the BMSCs of ICS group (P < 0.05). The ICSM group with ischemia seven days later showed better compared to the other two groups. The difference was significant ( P< 0. 05) . The brain iron content in the ICSM group was largely more than that in the ICS group (P<0.05). Conclusion External magnetic field can target delivery bone marrow mesenchymal stem cells labeled with superparamagnetic iron oxide in vitro and in vivo, and can obviously improve the transplantation of SPIO-Labeled mesenchymal stem cells.

Key words: Key words：magnetic field , bone marrow mesenchymal stem cells, superparamagnetic iron oxide , brain infarction, migration

中图分类号:

R743.32

郑洁陈晓丹郑希王顺和. 体外磁铁定向迁移SPIO标记的间充质干细胞对大鼠脑缺血治疗的影响[J]. 基础医学与临床, 2013, 33(10): 1302-1308.

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