Cell transplantation for the treatment of spinal cord injury in rats-morphological study and search for humoral effective factors-

细胞移植治疗大鼠脊髓损伤-形态学研究及体液有效因子的寻找-

基本信息

批准号：
20300122
负责人：
IDE Chizuka
金额：
$ 12.65万
依托单位：
Aino University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

项目摘要

(1) Transplantation of bone marrow stromal cells (BMSCs)(a) Direct injection of BMSCs into the spinal cord lesion BMSCs (5×10^5) were injected into the spinal cord lesion 2 weeks after injury, and rats were observed for 4 weeks post-transplantation. Locomotion of rats was recovered up to about 10 points of BBB scale, with the control points of 4-5. Numerous regenerating axons extended through the lesion from the rostral to distal border. Electron microscopy showed that regenerating axons were surrounded by Schwann cells in the collagen fibril matrices. BMSCs disappeared within 2 weeks post transplantation, indicating that they were never incorporated into the spinal cord.(b) Transplantation through the cerebrospinal fluid (CSF)BMSCs (5×10^5) were injected into the 4^<th> ventricle 1, 2, and 4 weeks after spinal cord injury. Locomotion of cell-injected rats was recovered up to 10-13 points of BBB scale, while those of control remained at 0-4. Regenerating axons extended in bundles bridg … More ing the lesion between the rostral and distal margin. Regenerating axons extended through the collagen fibril matrices and were surrounded by Schwann cells. This shows that regenerating axons are in the same environment as peripheral nerves. Injected BMSCs disappeared from the spinal cord 2 weeks after injection, indicating that BMSCs exert their effects not by integration into the spinal cord, but probably by secreting some trophic factors in to the CSF.(2) Effective factors contained in the conditioned medium of BMSCs It was demonstrated that the conditioned medium (CM) contained IGF-1, HGF, VEGF, and TGF-β1. Cultured hippocampal neurons had receptors of IGF-1 and TGF-β1. They showed no changes in density by the treatment of CM. These trophic factors, when they were used single or in combination of 2, 3 and 4 factors to treat cultured hippocampal neurons in vitro, did not show the same effect level of cell viability and neurite extension as the case of CM. This suggests that the CM may contain some unidentified trophic factors. Less

(1)骨髓基质细胞(BMSCs)移植(a)脊髓病灶内直接注射BMSCs 损伤后2周将BMSCs(5×10^5)注射到脊髓病灶内，观察大鼠4次。移植后几周，大鼠的运动能力恢复到 BBB 等级的约 10 点，控制点为 4-5 个再生轴突从损伤处延伸。电子显微镜显示再生轴突被胶原纤维基质中的雪旺细胞包围，在移植后两周内消失，表明它们从未并入脊髓。（b）通过脑脊液移植（脊髓损伤后1、2、4周将CSF)BMSCs (5×10^5)注射到第4脑室注射细胞的大鼠的运动恢复至 BBB 等级的 10-13 分，而对照组的再生轴突以束状延伸，桥接再生边缘。轴突延伸穿过胶原纤维基质并被雪旺细胞包围，这表明再生轴突与注射的周围神经处于相同的环境中。注射后2周，BMSCs从脊髓中消失，表明BMSCs发挥作用并非通过整合到脊髓中，而是可能通过向脑脊液分泌一些营养因子来发挥作用。 (2)BMSCs条件培养基中含有的有效因子结果表明，条件培养基（CM）含有IGF-1、HGF、VEGF和TGF-β1，培养的海马神经元的IGF-1和TGF-β1受体没有表现出变化。当这些营养因子单独或联合使用2、3和4种因子来处理体外培养的海马神经元时，没有表现出与CM处理相同的细胞活力和神经突延伸的效果水平。这表明 CM 可能含有一些未知的营养因子。