体外磁场定向迁移的间充质干细胞对脊髓损伤的治疗和GAP-43表达的影响.docVIP

体外磁场定向迁移的间充质干细胞对脊髓损伤的治疗和GAP-43表达的影响 郑 希，李成鹏，王顺和 (400016重庆，重庆医科大学：基础医学院病理学教研室1，干细胞与组织工程研究室2) [摘要] 目的 探讨体外磁场定向迁移尾静脉注射的SPIO标记的大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）对大鼠脊髓损伤治疗的影响以及对脊髓中GAP-43表达的影响。方法 体外培养SD大鼠BMSCs，流式细胞仪检测其免疫表型，超顺磁氧化铁颗粒（superparamagetic iron oxieds，SPIO）标记BMSCs，普鲁士蓝染色测定SPIO标记率，台盼蓝染色检测标记细胞存活率，CCK-8检测标记细胞分裂增殖能力，Western blot检测标记细胞干细胞标记物Nestin及GFAP表达。建立大鼠脊髓损伤（spinal cord injury，SCI）模型，尾静脉注射移植细胞，对细胞移植后大鼠进行BBB评分。普鲁士蓝染色观察脊髓组织内标记细胞，TTC染色观察脊髓缺血损伤面积，Western Blot检测损伤脊髓内GAP-43表达情况。结果 原代BMSCs体外能成功分离、培养、传代，且流式细胞仪检测表达神经干细胞标记物不表达造血干细胞标记物。普鲁士蓝染色显示SPIO标记率接近100%。台盼蓝染色显示标记细胞生存率与对照组无差异。CCK-8检测磁场下标记细胞增殖活性不受影响。Western blot检测磁场下标记细胞表达Nestin及GFAP无差异，分化活性不受影响。普鲁士蓝染色观察磁铁组脊髓组织中BMSCs明显多于无磁铁组。TTC染色显示磁铁组缺血面积小于无磁铁组。Western blot检测脊髓组织中同一时间点GAP-43表达，磁场组明显大于对照组及无磁场组。结论 外置磁场可以定向更多SPIO标记的BMSCs到达脊髓损伤处，使其更好地发挥修复作用且上调GAP-43 的表达，利于脊髓损伤的恢复。 [关键词] 磁场；骨髓间充质干细胞；超顺磁氧化铁；脊髓损伤；迁移；GAP-43 [中图法分类号] [文献标志码] A Effects of target delivery of superparamagnetic iron oxides-labeled bone marrow mesenchymal stem cells by a magnet on a rat model of spinal cord injury and the expression of GAP-43 preotein Zheng Xi, Li Chengpeng, Wang Shunhe (1Department. of Pathology, 2Laboratory of Stem Cell and Tissue Engineering, Institute of Basic Medical Sciences, Chongqing University of Medical Sciences,Chongqing 400016, China) [Abstract] Objective To investigate the effects of target delivery of magnet field on SPIO-labeled bone marrow mesenchymal stem cells（BMSCs） in vitro and observe whether the magnet field could promote labeled cells targeted migration and reduce the damage area of SCI in vivo.Methods BMSCs from Sprague Dawley(SD) were cultured in vitro. The expression of CD29、CD45、CD54 was checked by Flow cytometric detection. The BMSCs were labeled with SPIO and prussian blue staining was performed. The viability of labeled cells was evaluated by trypan blue staining. CCK-8 method was used to detect the proliferation of labeled cells with the presence magnetic field. Effects