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-

• 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
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20300122/
• 关键词: 神経再生; 移植・再生医療; 脊髄再生; 再生医学; 栄養因子; 骨髄間質細胞; 細胞移植; 神経再生; 移植・再生医療; 脊髄再生; 再生医学; 栄養因子; 骨髄間質細胞; 細胞移植

(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）将BMSC直接注射到脊髓病变BMSC（5×10^5）中2周后2周，将BMSC直接注射到脊髓病变中，并在传递后4周观察大鼠。将大鼠的运动恢复到BBB量表的约10点，控制点为4-5。许多再生轴突通过病变延伸从tho骨到远端边界。电子显微镜表明，再生轴突被胶原纤维材料中的雪旺细胞包围。 BMSC在移植后2周内消失，表明它们从未被掺入脊髓中。（b）通过脑脊液（CSF）BMSC（5×10^5）的移植被注入4^<th>心室1、2和4周后4周和4周。将注射大鼠的运动恢复高达10-13点BBB量表，而控制大鼠的刻度则保持在0-4。再生轴突延伸到捆绑式bridg中……更多的延髓和明显边缘之间的病变。再生轴突通过胶原原纤维材料延伸，并被雪旺细胞包围。这表明再生轴突与周围神经在相同的环境中。注射后2周从脊髓中消失了注射的BMSC，这表明BMSC不是通过将其整合到脊髓中而执行其效果，而是通过将一些营养因子分泌到CSF中。（2）BMSC条件介质中包含的有效因素，证明了条件介质（CM）包含IGF-1，HGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFCNe培养的海马神经元具有IGF-1和TGF-β1的受体。他们没有通过商业广告的处理显示密度变化。当这些营养因子被单一使用或组合2、3和4的组合来治疗体外治疗培养的海马神经元时，没有显示出与CM相同的细胞生存能力和神经蛋白神经蛋白扩展的效应水平。这表明CM可能包含一些未识别的营养因素。较少的