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.docVIP

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 　　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（P0.05）；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。 　　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（P0.05）.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 environm