骨髓间充质干细胞诱导成神经元样细胞移植治疗脊髓损伤  

Transplantation of neuron-like cells from bone marrow mesenchymal stem cells for treatment of spinal cord injury

作　　者：高平[1,2] 孙占胜[1] 王伯珉[1] 李连欣[1] 王甫[1] 牟乐明[1] 

机构地区：[1]山东大学附属省立医院创伤骨科,山东省济南市250021 [2]山东大学医学院,山东省济南市250012

出　　处：《中国组织工程研究》2013年第23期4256-4263,共8页Chinese Journal of Tissue Engineering Research

基　　金：山东省科技攻关计划项目(2006GG3202046)~~

摘　　要：背景:骨髓间充质干细胞诱导成为神经元样细胞移植治疗脊髓损伤已被证实有效,但不同诱导方法之间的差别尚无报道。目的:通过对脊髓损伤模型大鼠的行为对比及生化指标的检测,观察采用不同诱导方法将骨髓间充质干细胞诱导分化为神经元样细胞后移植对脊髓损伤疗效的差别。方法:取4周龄雄性Wistar大鼠骨髓分离培养骨髓间充质干细胞,对第3代骨髓间充质干细胞分别进行化学诱导和生物因子诱导后,收集备用。8周龄雄性Wistar大鼠48只,采用脊髓半横切法建立大鼠的脊髓损伤模型,随机分为4组:1周后骨髓间充质干细胞组损伤部位局部注射第3代骨髓间充质干细胞,化学诱导组局部注射化学诱导成的神经元样细胞,生物诱导组局部注射生物诱导成的神经元样细胞,DMEM培养液组局部注射细胞培养液。对48只大鼠脊髓损伤模型分别于伤后1,2,3,4,6,8,10,12周进行BBB评分,并于第12周末对损伤部位进行取材做组织切片,观察脊髓损伤的修复情况。结果与结论:模型建立后12周,骨髓间充质干细胞组、化学诱导组和生物诱导组大鼠后肢功能恢复明显优于DMEM培养液组(P<0.05),骨髓间充质干细胞组和化学诱导组功能恢复无明显差别(P=0.4363),生物诱导组恢复效果好于前2组(P<0.05)。生物诱导组大鼠运动功能持续恢复显著好于其他3组。脊髓组织切片苏木精-伊红染色显示骨髓间充质干细胞组和化学诱导组近似,脊髓胶质细胞增生,神经元样细胞崩解、空洞形成少于DMEM培养液组,生物诱导组神经损伤修复效果最佳。提示化学诱导后的骨髓间充质干细胞与未经过诱导的骨髓间充质干细胞在修复神经损伤的效果上没有明显差别;而经过生物诱导的骨髓间充质干细胞分化为神经元样细胞后移植治疗脊髓损伤的疗效明显优于未经诱导和化学诱导的骨髓间充质干细胞移植的疗效。BACKGROUND： The therapeutic effects of transplantation of neuron-like calls from bone marrow mesenchymal stem cells in the treatment of spinal cord injury have been confirmed. Howver, the efficiency differences among different methods of inducing neuron-like cell differentiation of bone marrow mesenchymal stem cells remain poorly understood. OBJECTIVE： After behavioral observation and biochemical index measurements of spinal cord injury rat models, the curative effects of transplantation of neuron-like cells induced by different methods from bone marrow mesenchymal stem calls in the treatment of spinal cord injury were investigated. METHODS： Bone marrow mesenchymal stem cells were isolated from 4-week-old male Wistar rats. Passage 3 bone marrow mesenchymal stem calls were induced by chemical methods and biological factors for later use. Spinal cord injury was induced in 48 male Wistar rats aged 8 weeks by spinal cord hemisection method. Then the rats were randomly divided into four groups： bone marrow mesenchymal stem cells, chemicaally-inducad, biological factor-induced and Dulbecco＇s modified Eagle＇s medium,（DMEM） groups, in which, passage 3 bone marrow mesenchymal stem calls, chemically-inducad or biological factor-inducad, or DMEM-treated bone marrow mesenchymal stem calls were injected into the injury region 1 week later, respectively. All 48 spinal cord injury rats were scored by the Basso, Beattie and Bresnahan locomotor rating scale at various time points （1, 2, 3, 4, 6, 8, 10, 12 weeks after injury）. At the end of 12 weeks after injury, tissue sections from the injury region were prepared for observation of repair of spinal cord injury. RESULTS AND CONCLUSION： At 12 weeks after spinal cord injury induction, recovery of hindlimb motor functions was superior in the bone marrow mesenchymal stem calls, chemically-induced, biological factor-induced groups to that in the DMEM group （P 〈 0.05）. There was no obvious difference in recovery of hindlimb motor function between bone marrow

关 键 词：干细胞 干细胞移植 脊髓损伤{细胞移植 诱导分化 生物诱导 化学诱导 省级基金 干细胞图片 文章 

分 类 号：R394.2[医药卫生—医学遗传学]