多次移植骨髓基质干细胞有利于脊髓损伤修复.docVIP

多次移植骨髓基质干细胞有利于脊髓损伤修复 作者：李辉映　文益民　 陈克明　蓝旭　汪大彬　孙振刚　胡凌云 作者单位：1.兰州大学第二临床医学院, 兰州 730018；2.兰州军区兰州总医院 a.脊柱外科;b.骨科研究所, 兰州 730050；　　基金项目：全军医学科学技术研究十一五计划课题项目（编号：06MA081） 　　【摘要】 ［目的］研究多次经蛛网膜下腔移植骨髓基质干细胞对Wistar大鼠脊髓损伤的功能修复作用。［方法］骨髓基质干细胞经体外分离、培养并用Hoecst33342标记。按照Allen方法把60只在T10-T12平面损伤的Wistar大鼠随机分4组，A、B、C、D组（对照组）。在损伤平面蛛网膜下腔中段放置一硅胶管，在7 d后，注入1×106个骨髓基质干细胞。在2、3、5、7、12周荧光显微镜、免疫组化检测骨髓基质干细胞在损伤段脊髓的存活、分布、分化情况并作计数观察。使用BBB评分观测后肢功能恢复。［结果］移植后7～14 d骨髓基质干细胞达到高峰，在7 d后表达巢蛋白及神经丝蛋白阳性。随时间的延长，移植在损伤部位的骨髓基质干细胞数量及神经元样细胞均有减少，但移植3次的细胞数减少速度较其他两组慢。移植3次组大鼠的BBB评分较移植1、2次组有明显的提高，P＜0.01,有统计学意义。［结论］多次移植骨髓基质干细胞更有利于脊髓损伤的恢复。 　　【关键词】 脊髓损伤　骨髓间充质干细胞 移植　蛛网膜下腔 　　 Repeated transplantation of bone marrow stromal cells into subarachnoid space to promote spinal cord injury repair 　　LI Huiying,WEN Yimin,CHEN Keming,et al. 　　The Second Clinic Hospital,Lanzhou University, Gansu 730018，China 　　Abstract：［Objective］To investigate the effect of multiple subarachnoid infusion of bone marrow stromal cells(BMSCs) on spinal cord injury repair in adult Wistar rats.［Method］The BMSCs were cultured, purified and labeled with Hoecst33342 in vitro. After the complete spinal cord injury rat model at T10T11 level was successfully established, sixty injured rats were randomly divided into 4 groups: groups A, B, C, and D. The labeled BMSCs in a dose of 1×106 /0.2 ml were injected into subarachnoid space of the rats via a catheters under the aubarachnoid space in group A,B and C. The cells were 　administered only once in group A 1week after injury. However, they were injected after injury 1 and 3 weeks in group B and 1, 3 and 5 weeks after injury in group C, respectively. Neurological functions were evaluated using the BassoBeattieBresnahan (BBB) Scale 1, 3，7，9 and 12 weeks after transplantation. The immunohistochemistry and fluorescence microscopy were analyzed to observe the migration, survival, differentiation and histomorphology of BMSCs. ［Result］The transplanted BMSCs survived and grew in the injured reg