神经干细胞移植对小鼠脊髓损伤的影响

The effect of neural stem cells transplantation on spinal cord injury in mice

目的观察神经干细胞(NSCs)移植对脊髓损伤(SCI)小鼠功能恢复的影响。方法分离、培养、增殖和纯化E14-17d小鼠的NSCs,并通过免疫荧光法对其进行鉴定。将绿色荧光蛋白转基因小鼠的NSCs移植到小鼠脊髓损伤模型体内,术后进行行为学和病理学检测,观察小鼠功能恢复情况。运用改良Allen's法制备小鼠T10-T11脊髓损伤动物模型,动物分为假手术组(12只)、模型组(12只),治疗组(12只)和对照组6(12只)。治疗组每只小鼠自眶静脉注射NSCs悬液200μl(2×10个细胞),对照组只注射DMEM/F12培养基200μl。术后1d、3d、7d、14d、21d、28d和56d,进行BBB运动功能评分和病理学检测,观察植入区细胞生长变化情况。结果 BBB评分显示:治疗组明显高于对照组(<0.01),治疗组与假手术组相比没有明显差异(>0.05),说明NSCs移植后小鼠的行为学得到了明显改善,功能有所恢复。病理学检测发现,移植后NSCs不仅迁移到脊髓损伤区,而且与宿主细胞较好地整合。结论移植的E14-17d胚胎小鼠NSCs不仅可在脊髓损伤部位存活并和宿主细胞整合,而且可促进小鼠后肢运动功能恢复。

Objective To observe the effects of neural stem cells（NSCs） transplantation on the function recovery of mouse after spinal cord injury.Methods NSCs from E14-17 days mouse were isolated,cultured,proliferated,purified and identified by immunofluorescence.The NSCs from E14-17 days green fluorescent protein transgenic mouse were transplanted into the animal model of spinal cord injury.The functional recovery of mouse was assessed by the behavioral and pathological examination after cells transplantation.The model of spinal cord injury from T10 to T11 mice was established with Allen＇s method in which we made a little adjustment.Mice were divided into sham operational group（12）,model group（12）,transplantation group（12） and control group（12）.200μl of NSCs suspension（2×106cells） was grafted into mice by orbital intravenous injection in transplantation group,and 200μl of DMEM/F12 was injected into mice in control group.1d,3d,7d,14d,21d,28d and 56d after operation,the BBB score and pathological motor function was determined,and the growth of being grafted cells was observed.Result BBB scores were significantly higher in transplantation group than in control（0.01）,but without significant differrence between transplantation group and sham group（0.05）,which indicated that transplantation of NSCs improved the function of spinal cord injury.Pathological examination proved that NSCs not only migrated to injury zone,but also surrounded injury zone and made a better integration with the host cells.Conclusion The transplantation of NSCs from E14-17days mice can not only survive and integrate with host cells,but also improve the funcion of the spinal cord injury mice.