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人脐带间充质干细胞与自体激活雪旺细胞联合移植修复脊髓损伤的实验研究
引用本文:朱玉海,冯世庆,孔晓红,王雪,孟恒星,李伟,李健辉. 人脐带间充质干细胞与自体激活雪旺细胞联合移植修复脊髓损伤的实验研究[J]. 中华创伤骨科杂志, 2009, 11(8). DOI: 10.3760/cma.j.issn.1671-7600.2009.08.012
作者姓名:朱玉海  冯世庆  孔晓红  王雪  孟恒星  李伟  李健辉
作者单位:天津医科大学总医院骨科,300052
基金项目:教育部新世纪优秀人才支持计划,天津市应用基础和前沿技术支持计划项目,天津市卫生局科研基金 
摘    要:目的 通过观察人脐带间充质干细胞(hUCMSCs)和大鼠自体激活雪旺细胞(AASCs)联合移植修复脊髓损伤的疗效,探讨AASCs对hUCMSCs体内存活、分化的影响.方法 分离、培养hUCMSCs和大鼠AASCs.通过IMPACTOR MODEL-Ⅱ型打击仪将80只Wistar成年雌性大鼠均制作成T10损伤模型,随机分为四组(n=20):DMEM移植对照组、hUCMSCs移植组、AASCs移植组、hUCMSCs与AASCs联合移植组.比较符组动物恢复情况,进行行为学评分(BBB评分),NF-200和GFAP染色观察细胞存活、分化情况,生物素葡聚糖胺示踪观察皮质脊髓束再生情况.结果 4周后各组间BBB评分差异有统计学意义(P<0.05),6周后联合移植组明显高于其他三组,差异有统计学意义(P<0.05).免疫组化染色示联合移植组的hUCMSCs存活数量,NF-200、GFAP阳性荧光面积均明显高于hUCMSCs移植组,差异有统计学意义(P<0.05),BDA顺行爪踪可见联合移植组于损伤区染色较多,部分纤维延续至损伤远端.结论 AASCs可支持移植的hUCMSCs在损伤部位存活并向神经方向分化,hUCMSCs与AASCs联合移植较二者单独移植能更有效地促进脊髓损伤后运动功能的恢复和轴突再生.

关 键 词:脊髓损伤  细胞移植  神经再生  脐带间充质干细胞  雪旺细胞

Repair of spinal cord injury with co-transplantation of human umbilical cord mesenchymal stem cells and autologously activated Schwann cells
Abstract:Objective To observe the effect of co-transplantation of human umbilical cord mes-enchymal stem cells (hUCMSCs) and autnlognusly activated Schwann cells (AASCs) on the recovery of acute spinal cord injury (SCI),as well as the influence of AASCs on the survival and differentiation of hUCMSCs. Methods hUCMSCs were isolated from human umbilical cord,cultured and purified in vitro. Unilateral saphenous nerves were ligated to isolate AASCs. Eighty female Wistar rats [(77±2) d] were randomly as-signed to 4 even study groups: DMEM control,hUCMSCs transplantation,AASCs transplantation;and co-transplantation. The acute spinal cord contusion models (T10) were established by IMPACTOR MODEL-Ⅱ device. Recovery of the lower extremity was observed using Basso- Beattie- Bresnaban (BBB) locomotor scoring system,and immunohistnchemical stain to detect the survival and differentiation of hUCMSCs. Re-generation of the corticospinal tract (CST) was observed using the biotinylated dextran amine (BDA) tracing. Results The BBB scores in cell transplantation groups were significantly higher than in the DMEM control group 4 weeks after injury (P<0.05 ) . The co-transplantation group had a higher score than the hUCMSCs group(P<0.05) . Both the number of surviving hUCMSCs and the positive response area in im-munohistochemistry staining were significantly larger in the co-transplantation group than in other groups 6 weeks after injury(P<0.05) . The BDA anterograde tracing showed more regenerating nerve fibers got through the injured area in the co-transplantation group which also had the smallest injured cavity,compared with other groups. Conclusions AASCs can promote survival of hUCMSCs and their differentiation into neural cells. Co-transplantation of hUCMSCs and AASCs may be more effective in promoting functional re-covery and axonal regeneration after SCI.
Keywords:Spinal cord injury  Cell transplantation  Neural regeneration  Umbilical cord mesenchymal stem cells  Schwann cells
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