细胞联合移植修复大鼠脊髓损伤的可行性  

Possibility of cell co-transplantation for rats with spinal cord injury

作　　者：吴立生[1] 吴仕峰[1] 

机构地区：[1]临沂市人民医院创伤二科,山东省临沂市276000

出　　处：《中国组织工程研究与临床康复》2008年第34期6626-6630,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景：嗅鞘细胞和骨髓间充质干细胞均可修复脊髓损伤，两者联合能否发挥出更佳的修复效果还不太清楚。目的：观察唉鞘细胞和骨髓间充质干细胞联合移植修复脊髓损伤的效果。设计、时间及地点：随机对照动物实验，于2006-06／2007-10在临沂市人民医院和昆明医学院完成。材料：健康清洁级成年SD大鼠，雌性，体质量220～280g，分为：正常对照组（n=5）、嗅鞘细胞移植组（n=10）、骨髓间充质干细胞移植组（n=10）、联合细胞移植组（n=10）、手术对照组（n=10）。方法：分离大鼠嗅球组织和股骨，分离培养嗅鞘细胞和骨髓间充质干细胞。暴露大鼠脊髓予以尖刀离断制备完全脊髓损伤动物模型，正常对照组仅打开椎板未行脊髓离断，嗅鞘细胞移植组、骨髓间充质干细胞移植组、联合移植组分别在断端及远近端注射嗅鞘细胞、骨髓间充质干细胞及嗅鞘细胞＋骨髓间充质干细胞悬液，采取3点注射，3点的注射深度分别为0．5mm、1．0mm和1．5mm。手术对照组在脊髓离断后仅注射培养基。主要观察指标：将嗅鞘细胞和骨髓间充质干细胞制成细胞悬液后接种培养，并应用P75抗体和BrdU抗体对联合培养的细胞进行免疫荧光鉴定。移植后评定大鼠脊髓功能恢复情况，并观察病理学改变和细胞存活情况。结果：联合培养的细胞部分相互编织成网，部分呈条索状，荧光鉴定双标阳性者为嗅鞘细胞，只发红色荧光而不发绿色荧光者为骨髓间充质干细胞。移植嗅鞘细胞和骨髓间充质干细胞的大鼠脊髓功能均有改善，移植4周后，联合移植组脊髓功能恢复优于单细胞移植组。病理学观察显示，手术对照组脊髓结构破坏严重，损伤处可见大量空洞及神经元胞体萎缩，周围存在较多的空洞及组织液化，细胞数目较少，神经纤维排列紊乱；移植鞘细胞及骨髓间充质干细�BACKGROUND： Olfactory ensheathing cells （OECs） and bone marrow mesenchymal stem cells （BMSCs） repair the function of injured spine cord. Whether their combination can have better repair effect is unclear. OBJECTIVE： To investigate effects of OEC and BMSC transplantation for treating injured spine cord. DESIGN, TIME AND SETTING： The randomized control animal experiment was performed at the Linyi People＇ s Hospital and Kunming Medical College from June 2006 to October 2007. MATERIALS： Healthy clean adult female Sprague Dawley rats with the body mass of 220-280 g were enrolled and divided into a normal control group （n=5）, a OEC group （n=10）, a BMSC group （n=10）, a co-transplantation group （n=10） and a operation control group （n= 10）. METHODS： OECs and BMSCs were harvested from the olfactory bulb and femoral bone. Animal models of complete spinal cord injury were established by cutting rat spinal cord. Rats in the normal control group only received vertebral plate unfolding, but no spinal cord transaction. Rats in the OEC group, BMSC group and co-transplantation group were respectively injected with OECs, BMSCs and their combination at the broken ends and proximal and distal ends. Using three points injection, the injection depth was separately 0.5 mm, 1.0 mm and 1.5 mm. Rats in the operation control group only received medium following spinal cord transaction. MAIN OUTCOME MEASURES： OECs and MSCs were co-cultured after being digested. Cells were identified by P75 and BrdU immunofluorenscence. Spinal cord functional recovery was assessed, and pathological changes and cell survival were observed after transplantation. RESULTS： One proportion of co-cultured cells wove into net with their processes linking. The other proportion grew like strip-shaped. OECs were identified on the cell membrane （green fluorescence） and on the cell core （red fluorescence）, whereas BMSCs were identified on the cell core （red fluorescence） without cell membrane （green fluorescence）

关 键 词：嗅鞘细胞 骨髓间充质干细胞 脊髓损伤 细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]