长期培养人胚神经干细胞对兔脊髓损伤的修复作用  

Repairing effect of human neural stem cells (hNSCs) on rabbit spinal cord injury

作　　者：阳运康[1] 林月秋[2] 汤逊[1] 蔡培强[1] 

机构地区：[1]成都军区昆明总医院 [2]成都军区昆明总医院骨科,云南昆明650032

出　　处：《中国矫形外科杂志》2005年第16期1244-1247,共4页Orthopedic Journal of China

基　　金：云南省自然科学基金资助项目(2002C0070M)

摘　　要：[目的]观察长期培养人胚神经干细胞(hNSCs)的体外生长特性与转染EGFP基因后移植治疗兔脊髓横切损伤模型在体内的生物学活性及对神经结构修复和功能恢复的影响。[方法]体外分离、培养并鉴定hNSCs,用逆转录病毒介导的增强绿色荧光蛋白基因(EGFP)进行转染;制备兔T9全横断脊髓损伤模型;观察hNSCs移植对脊髓损伤后神经结构修复和功能恢复的影响。[结果]从胎龄10～20周的新鲜人胚脑皮层中成功分离出神经干细胞,该细胞具有连续克隆传代能力,诱导分化后表达分化细胞的特异抗原。本实验室已成功连续培养10个月(17代),转染EGFP基因后,仍保持未分化状态,能够自我更新形成新的神经球;移植入兔SCI模型后,hNSCs能在体内存活、迁移、分化并增殖。与对照组相比,hNSCs移植组明显促进了脊髓神经的再生、结构的修复和下肢运动功能的恢复。[结论]hNSCs移植促进了脊髓损伤后神经结构的修复和功能的恢复,是治疗急性脊髓损伤的一种有效方法。[ Objective ] o investigate the growth characterization, of human neural stem cells （ hNSCs ） while long-timely cultured or transfected with retrovial-mediated EGFP gene in vitro, the biological activities and effects on repairing neural morphology and recovering function after transplanted to the models of spinal cord injury in rabbits were recorded and analyzed. [Method] Neural stem cells from fresh embryonic cortex following abortion were isolated, cultured, identificated and transfected with retrovial-mediated EGFP gene in vitro. Transection of spinal cord at the level of I9 was produced at 32 adult rabbits. Neural morphological repairing and functional recovery were investigated after hNSCs was grafted into the injury site of spinal cord. [Result] Human neural stem cell line from fresh embryonic cortex in El0 to E20 weeks could be cloned and passaged continuously. Mature neural cells after differentiation could express specific antigen. It had been passaged continuously for 10 months（ 17 passages） keeping strengthened capacity of proliferation in labg. These hNSCs can be genetically engineered to express foreign transgenes in vitro. It remained undifferentiated and has self-renewing capacity and potential multipotency of differentiation after transfected with EGFP gene. After transplanted to the adult rabbits models of spinal cord injury, it were viable, migratory, proliferous, as well as improving neural regeneration, morphological repairing and functional recovery compared to the control. [ Conclusion] Transplantation of neural stem cells could improve neural morphological repairing and functional recovery abilities on spinal cord injury. Neural stem cells transplantation is an effective method for treatment of spinal cord injury.

关 键 词：人胚神经干细胞 脊髓损伤 移植 转染 急性脊髓损伤 修复作用 长期培养 绿色荧光蛋白基因 EGFP基因 逆转录病毒介导 

分 类 号：R651.2[医药卫生—外科学]