神经生物膜介导神经干细胞联合雪旺细胞移植治疗脊髓损伤  

Treatment of spinal cord injury by neurobiological membrane guided neural Stem cells combined with schwann cells transplantation

作　　者：冯世庆[1] 王金宝[1] 孔晓红[2] 陈家童[2] 纪江峰[1] 荆峰[1] 周先虎[1] 侯巍[1] 

机构地区：[1]天津医科大学总医院骨科,300052 [2]南开大学生命科学院

出　　处：《中华实验外科杂志》2006年第11期1401-1403,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(30271313)

摘　　要：目的探讨以壳聚糖-胶原偶联生物膜为载体,联合移植雪旺细胞(SCs)与神经干细胞(NSCs)在治疗脊髓损伤中的作用,并评价该方法的疗效。方法大鼠孕鼠体内取出胎鼠分离获得原代NSCs进行体外培养,大鼠乳鼠坐骨神经中分离获得原代SCs进行体外培养,分别传代4代获得大量细胞。40只Wistar大鼠建立大鼠脊髓半横断动物模型并随机分为4组：空白对照组,SCs移植组,NSCs移植组,NSCs联合SCs移植组。将胎鼠NSCs和乳鼠SCs共同种植于胶原-壳聚糖偶联的神经生物膜上,移植入大鼠脊髓半横断模型的脊髓损伤部位,对动物进行BBB脊髓功能评分,BDA神经示踪标记,标记2周后处死动物取出动物脊髓组织进行Cy3荧光素染色,p75免疫荧光染色及脊髓组织苏木素-伊红(HE)染色。结果4组实验动物BBB评分组间比较差异有统计学意义(ANOVA,P＜0．05),p75免疫荧光染色阳性物面积比较各组之间差异有统计学意义(ANOVA,P＜0．05)。结论NSCs和SCs能够在胶原．壳聚糖偶联的神经生物膜上共同生长分化,并且SCs能够诱导NSCs产生轴突定向生长。壳聚糖-胶原生物膜介导的SCs联和NSCs移植治疗脊髓损伤可使神经部分再通。Objective To observe the effect of transplantation of Schwann cells and neural stem cells embedded to the chitosan-collagen complex film to the spinal cord injury site in Wistar rats. Methods Schwann cells from the sciatic nerve of the Wistar rats were isolated and purified. A neural stem cell line which had ability to form clones was isolated from the brain of Wistar rat embryo. The clone was passaged continuously for 4 generations and large number of clones obtained. The two type cells were embedded to the chitesan-collagen complex film and the compatibility analyzed between them. Then the 40 adult Wistar rats, weighing （200 ±30） g, were divided into 4 groups： Blank group, SCs groups, NSCs group and compound cells group and all rats were hemi-sectioned on right spinal cord （T10）. Then these films were transplanted in the spinal cord injury site. Forteen weeks later, two rats in each group selected at random were injected with 10 % BDA in the double cerebra cortex for anterograde labeling of the spinal cord. After 2 weeks, these rats were killed and poured with 4 % PLA. The spinal cords including the segment of defect were taken out and cut into 5μm thick sections at 0 ℃. The sections were stained by cy3, HE, and immunohistochemistry was used to detect p75. Results The BBB scores in cells groups were higher than in the blank group. The recovery of motion function in cells groups was quicker than in the blank group （ANOVA,P 〈 0.05）. The regenerated nerve fiber stained by cy3 and HE could be seen in the defected position in cells group and blank group, almost nothing in the injured site. There was significant difference between cells group and blank group （ANOVA, P 〈 0.05）. Conclusion The two types of cells have good compatibility with the chitosan-collagen complex film. Transplantation of Schwann cells and neural stem cells embedded to the chitosan-collagen complex film can guide regeneration.

关 键 词：神经干细胞 雪旺细胞 脊髓损伤 胶原 

分 类 号：R651.2[医药卫生—外科学]