人胚嗅鞘细胞与鼠胚胎脊髓组织联合移植对大鼠脊髓损伤的治疗(英文)  

Combined transplantation of human fetal olfactory ensheathing cells and rat embryonic spinal cord tissues in the treatment of spinal cord injury in rats

作　　者：吴卫江 惠国桢[2] 吕然博[3] 苗宗宁 

机构地区：[1]无锡第三人民医院细胞室,江苏省无锡市214000 [2]苏州大学附属第一医院神经外科,江苏省苏州市215000 [3]东南大学附属中大医院神经外科,江苏省南京市210009

出　　处：《中国临床康复》2006年第41期175-179,I0006,共6页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然基金项目(30271325);江苏省自然基金资助项目(BK2001170)~~

摘　　要：背景:脊髓损伤高发且后果严重,至今尚无有效措施挽救丧失的神经功能。哺乳动物嗅觉系统的一类特殊胶质细胞的移植引起关注。目的:观察人胚嗅鞘细胞和大鼠胚胎脊髓共同移植在促进大鼠横断脊髓轴索再生方面是否产生协同作用。设计:开放性实验。单位:无锡第三人民医院细胞室,苏州大学附属第一医院神经外科,东南大学附属中大医院神经外科。材料:实验于2002-09/2004-10在无锡市第三人民医院细胞实验室完成。①选取清洁级成年雌性SD大鼠36只,随机数字表法分成4组:人胚嗅鞘细胞组10只、鼠胚胎脊髓组10只、联合移植组10只,模型组6只。②取12周左右的流产新鲜人胚胎(产妇知情同意)用于嗅鞘细胞的培养纯化。③取孕14d的SD大鼠1只,施行剖腹手术将胎鼠连同胎膜一同取出,用于新鲜胚胎脊髓的制备。方法:①4组大鼠均建立脊髓半切洞模型。模型组在损伤洞腔内填塞明胶海绵加全培养基8μL,在损伤上下各1mm处注射相同培养基2μL;人胚嗅鞘细胞组明胶上给予嗅鞘细胞悬液8μL,损伤上下各1mm处注射细胞悬液2μL;鼠胚胎脊髓组将组织碎块直接填塞在洞腔内,外覆明胶;联合移植组将同样大小的胚胎脊髓填塞在洞腔内,然后用微量加样器将8μL的嗅鞘细胞悬液注入洞腔,外覆明胶,在洞腔上下各1mm处注射细胞悬液2μL,按层缝合肌层皮肤。②定期对各组大鼠进行行为学评定,结合病理学观察,并通过辣根过氧化物酶-四甲基联苯胺逆行示踪技术,评价嗅鞘细胞和胚胎脊髓对神经元存活、纤维再生的影响。主要观察指标:①人胚嗅鞘细胞体外培养及纯化。②体外免疫细胞化学分析。③大鼠后肢运动功能BBB评分测定结果。④移植物及受损脊髓修复的免疫组化检测结果。⑤辣根过氧化物酶-四甲基联苯胺示踪对各组被标记的皮质及中脑红核神经元的定量分析情况。结果:①人�BACKGROUND： Spinal cord injury occurs frequently and its consequence is very. severe. There is no effective method to rebuild the function of demylinated nerves. Transplantation of a kind of special glial cells in olfactory system of mammal attracts more attention. OBJECTIVE： To observe whether combined transplantation of human fetal olfactory ensheathing cells （human OECs） and rat embryonic spinal cord tissues （rat ECS） possesses synergistic effect in promoting axonal regeneration in the rats following spinal cord transection. transection. DESIGN： Open experiment. SETTING： Cell Room, Third People＇s Hospital of Wuxi City; Department of Neurology, First Hospital Affiliated to Soochow University; Department of Neurosurgery, Zhongda Hospital Affiliated to Central South University MATERIALS： This experiment was carried out in the cell laboratory of the Third People＇s Hospital of Wuxi from September 2002 to October 2004. （1） Totally 36 adult female SD rats, of clean grade, were selected and randomly divided into 4 groups： human OECs group （n=10）, rat ECS group （n=10）, combined transplantation group （n=10） and sham-operation group （n=6）. （2） Fresh 12-week aborted human embryo was used for culture and purification of human OECs （Informed consent was obtained from the parturient）. （3）One SD rat at embryonic 14 days underwent caesarean operation, and fetal rat and fetal membrane were taken out together and used for preparing new embryonic spinal cord. METHODS： （1）Rats of 4 groups were all created into hemiseetion cavity models. Gelatin sponge and complete culture medium of 8 μL were packed into the injured cavity of rats in the model group, and the same culture medium of 2 μL was injected at 1 mm above or below injure; Human OECs suspension of 8 μL was added to gelatin sponge in human fetal Human OECs group, and human OECs suspension of 2 μL was injected at 1 mm above and below injure; rat embryonic spinal cord tissue of rat ECS group was chipped

关 键 词：细胞移植 嗅神经 脊髓/胚胎学 脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]