间充质干细胞与神经干细胞序贯移植修复大鼠脊髓损伤的作用研究(1)
摘要：目的 通过比较由单独神经干细胞(NSCs)移植治疗大鼠脊髓损伤(SCI)和由骨髓间充质干细胞(BMSCs)与NSCs序贯治疗大鼠SCI的脊髓恢复情况不同，研究BMSCs对大鼠脊髓微环境的影响以及序贯治疗的可行性和优势。方法 体外分离，培养，纯化NSCs和BMSCs，改良Allen法制造大鼠脊髓损伤(SCI)模型，48只大鼠随机分为单独标记神经干细胞移植组(A组，n=24)，间充质干细胞+标记神经干细胞序贯移植组(B组，n=24)。分别于移植后3 d、1 w、2 w，3 w、4 w、6 w、8 w取脊髓组织，观察脊髓空洞形成情况，神经干细胞分化生长情况，以及脊髓胶质疤痕大小。结果 序贯移植组BBB评分比单独神经干细胞移植组高(P<0.05)；观察大鼠在治疗后1，2，3，4，6，8 w时的神经元细胞数目，血管生长情况，胶质疤痕以及脊髓空洞大小A，B组之间有显著性差异(P≤0.05)。结论 BMSCs可促进了NSCs的有效分化，减少胶质化倾向，改善了脊髓的微环境有利于治疗SCI，骨髓间充质干细胞与神经干细胞序贯移植对大鼠脊髓损伤的疗效优于单独移植神经干细胞。
, 百拇医药
关键词：脊髓损伤；细胞培养；移植；慢病毒转染；序贯移植
中图分类号：R651.2 文献标识码：A 文章编号：1006-1959(2017)20-0038-05
Abstract：Objective To compare by separate neural stem cells(NSCs)transplantation for the treatment of spinal cord injury(SCI)and bone marrow mesenchymal stem cells(BMSCs)and NSCs sequential therapy of SCI rats spinal cord recovery，effects of BMSCs on rat spinal cord microenvironment and sequential treatment of culture method of feasibility and advantage.Methods In vitro，separation，purification of NSCs and BMSCs in rats with spinal cord injury，manufacturing method of modified Allen(SCI)model，48 rats were randomly divided into separate labeled neural stem cells transplantation group(group A，n=24)，Mesenchymal stem cells+labeled neural stem cells in the sequential transplantation group (group B，n=24).The spinal cord tissue was observed at 3 d，1 w，2 w，3 w，4 w，6 w and 8 w after transplantation.The formation of spinal cord formation， the growth of neural stem cells and the size of glial scar were observed.Results The BBB score of the sequential transplantation group was higher than that of the neural stem cell transplantation group(P<0.05).The number of neuronal cells，the growth of the blood vessels were observed at 1，2，3，4，6 and 8 w after treatment.There was a significant difference between the quality scar and the size of the syringia in group A and B(P≤0.05).Conclusion BMSCs can promote the differentiation of NSCs effectively，reduce glial tendency，improve the micro environment of the spinal cord is conducive to the treatment of SCI，bone marrow mesenchymal stem cells and neural stem cells transplantation on curative effect is better than that of sequential injury of rat spinal cord neural stem cells transplantation alone.
, 百拇医药
Key words：Spinal cord injury；Cell culture；Transplantation；Lentivirus transfection；Sequential transplantation
脊髓損伤(spinal cord injury，SCI)可导致严重的运动、感觉和自主神经功能障碍，它是一种高花费、高致残率的疾病，目前，对受伤的脊髓还没有有效的治疗方法[1]。脊髓损伤的治疗一直是骨科医生的研究热点和重点，然而手术只能解除对脊髓的压迫和恢复脊柱的稳定性，还无法使损伤的脊髓恢复功能[2]，而应用SCs的多向分化潜能，可生成各种组织细胞，特别是特定的神经细胞，这使得修复损伤的脊髓组织，恢复脊髓功能成为可能，应用SCs移植有可能治愈SCI，是最具潜力的治疗方法。近年来多项研究已经发现神经干细胞和骨髓间充质干细胞局部移植均可促进脊髓损伤的修复[3-4]，但深入研究发现：移植的NSCs在体内存活较少，且即使存活分化也存在困难，原因是脊髓损伤后微环境不利于NSCs的存活，定植及分化。在2013年，Forostyak提出了间充质干细胞能改善脊髓损伤的微环境[5]。同时我们体外实验发现：MSCs促进了NSCs的有效分化[6-7]。那么，其在体内是否显示同样的效应？为了探明在动物体内间充质干细胞是否对脊髓微环境起改善作用以及间充质干细胞对神经干细胞分化的影响，探索序贯移植的有效性及其方法，我们利用大鼠的脊髓损伤模型分别进行了序贯移植与单独神经干细胞移植治疗大鼠SCI的动物实验。, 百拇医药(陈志 申才良)
, 百拇医药
关键词：脊髓损伤；细胞培养；移植；慢病毒转染；序贯移植
中图分类号：R651.2 文献标识码：A 文章编号：1006-1959(2017)20-0038-05
Abstract：Objective To compare by separate neural stem cells(NSCs)transplantation for the treatment of spinal cord injury(SCI)and bone marrow mesenchymal stem cells(BMSCs)and NSCs sequential therapy of SCI rats spinal cord recovery，effects of BMSCs on rat spinal cord microenvironment and sequential treatment of culture method of feasibility and advantage.Methods In vitro，separation，purification of NSCs and BMSCs in rats with spinal cord injury，manufacturing method of modified Allen(SCI)model，48 rats were randomly divided into separate labeled neural stem cells transplantation group(group A，n=24)，Mesenchymal stem cells+labeled neural stem cells in the sequential transplantation group (group B，n=24).The spinal cord tissue was observed at 3 d，1 w，2 w，3 w，4 w，6 w and 8 w after transplantation.The formation of spinal cord formation， the growth of neural stem cells and the size of glial scar were observed.Results The BBB score of the sequential transplantation group was higher than that of the neural stem cell transplantation group(P<0.05).The number of neuronal cells，the growth of the blood vessels were observed at 1，2，3，4，6 and 8 w after treatment.There was a significant difference between the quality scar and the size of the syringia in group A and B(P≤0.05).Conclusion BMSCs can promote the differentiation of NSCs effectively，reduce glial tendency，improve the micro environment of the spinal cord is conducive to the treatment of SCI，bone marrow mesenchymal stem cells and neural stem cells transplantation on curative effect is better than that of sequential injury of rat spinal cord neural stem cells transplantation alone.
, 百拇医药
Key words：Spinal cord injury；Cell culture；Transplantation；Lentivirus transfection；Sequential transplantation
脊髓損伤(spinal cord injury，SCI)可导致严重的运动、感觉和自主神经功能障碍，它是一种高花费、高致残率的疾病，目前，对受伤的脊髓还没有有效的治疗方法[1]。脊髓损伤的治疗一直是骨科医生的研究热点和重点，然而手术只能解除对脊髓的压迫和恢复脊柱的稳定性，还无法使损伤的脊髓恢复功能[2]，而应用SCs的多向分化潜能，可生成各种组织细胞，特别是特定的神经细胞，这使得修复损伤的脊髓组织，恢复脊髓功能成为可能，应用SCs移植有可能治愈SCI，是最具潜力的治疗方法。近年来多项研究已经发现神经干细胞和骨髓间充质干细胞局部移植均可促进脊髓损伤的修复[3-4]，但深入研究发现：移植的NSCs在体内存活较少，且即使存活分化也存在困难，原因是脊髓损伤后微环境不利于NSCs的存活，定植及分化。在2013年，Forostyak提出了间充质干细胞能改善脊髓损伤的微环境[5]。同时我们体外实验发现：MSCs促进了NSCs的有效分化[6-7]。那么，其在体内是否显示同样的效应？为了探明在动物体内间充质干细胞是否对脊髓微环境起改善作用以及间充质干细胞对神经干细胞分化的影响，探索序贯移植的有效性及其方法，我们利用大鼠的脊髓损伤模型分别进行了序贯移植与单独神经干细胞移植治疗大鼠SCI的动物实验。, 百拇医药(陈志 申才良)
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