成年脊髓损伤大鼠脊髓神经干细胞的体外培养及分化研究  

Culture and differentiation of neural stem cells from the injured spinal cord in adult rats

作　　者：叶正旭[1] 李静[2] 黄景辉[1] 闫铭[1] 梁伟[1] 杨旻[1] 罗卓荆[1] 

机构地区：[1]第四军医大学西京医院骨科,西安710032 [2]第四军医大学基础部病理学教研室

出　　处：《中华创伤骨科杂志》2009年第10期956-959,共4页Chinese Journal of Orthopaedic Trauma

基　　金：基金项目：国家高技术研究发展计划（863计划）资助项目（2002AA216101）;国家教育部骨干教师资助计划项目

摘　　要：目的探讨大鼠脊髓损伤后脊髓神经干细胞的分离培养方法及分化情况。方法采用Allen法制作大鼠脊髓损伤模型，利用无血清培养和单细胞克隆技术在成年脊髓损伤7d大鼠脊髓中分离具有单细胞克隆能力的神经干细胞，并进行培养鉴定。结果从成年脊髓损伤7d大鼠脊髓中成功分离出神经干细胞，该细胞具有连续克隆能力，可传代培养，表达神经巢蛋白抗原。分化后的细胞表达神经元细胞、星形胶质细胞和少突胶质细胞的特异性抗原。结论致伤7d的成年大鼠脊髓组织体外可培养出神经干细胞，并分化为神经元细胞、星形胶质细胞和少突胶质细胞，有可能参与脊髓损伤的修复过程。Objective To investigate culture and differentiation of neural stem cells from the injured spinal cord in adult rats. Methods The spinal cord of male SD rats was injured by Allen＇s weight dropping on T8. Procedures of floating neural stem cell culture were carried out. Results Neurospheres of neural stem cells emerged after plating for 1 week. They had potent self-renewing ability and could be passaged repeatedly. Immunohistochemistry data showed that these neurospheres strongly expressed nestin. A characteristic intermediate filament was seen in neural stem cells. The stem cells were further induced to differentiate into cells expressing β-tubulin, glial fibrillary acidic protein （GFAP） and myelin associated glycoprotein （MAG）, suggesting their differentiating potencies into neurons, astrocytes and oligodendrocytes. Conclusion In adult rats after spinal cord injury the neural stern cells can be isolated and cultured in vitro and induced to differentiate into neurons, astrocytes and oligodendrocytes, so the neural stem cells may participate in repair of spinal cord injury.

关 键 词：脊髓 脊髓损伤 干细胞 细胞分化 大鼠 

分 类 号：R686[医药卫生—骨科学]