绿色荧光蛋白转基因大鼠骨髓间充质干细胞在急性脊髓损伤大鼠中的迁移和分化

Migration and differentiation of bone mesenchymal stem cells from green fluorescent protein transgenic rats with spinal cord injury

目的观察绿色荧光蛋白转基因大鼠来源的骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在急性脊髓损伤大鼠脊髓组织中的迁移和分化情况。方法全骨髓贴壁培养法培养BMSCs,Allen法制作脊髓损伤大鼠模型,共30只大鼠。于造模1周后进行干预,随机选取造模成功的24只大鼠并随机分为脊髓损伤组、假移植组(生理盐水注射)和细胞移植组(BMSCs注射),每组8只。于移植前、移植后7 d、14 d、21 d、28 d、35 d及42 d进行BBB(Basso,Beattie&Bresnahan locomotor rating scale)评分。HE染色、免疫荧光观察脊髓损伤的组织修复和BMSCs的迁移分化情况。结果从第14天始,细胞移植组大鼠的BBB评分较脊髓损伤组和假移植组大鼠高,差异具有统计学意义(P<0.05)。HE染色显示细胞移植组大鼠的脊髓结构相对完整,液化和囊泡区缩小,炎性细胞减少。免疫荧光显示BMSCs聚集于脊髓损伤处,且能分化为神经元、神经胶质细胞和神经前体细胞。结论 BMSCs能向脊髓损伤部位迁移和聚集,并分化为相应的神经细胞促进脊髓功能恢复。

Objective To investigate migration and differentiation of bone mesenchymal stem cells（ BMSCs） from green fluorescent protein（ GFP） transgenic rats with spinal cord injury.Methods BMSCs were harvested by whole bone marrow adherent culture method,and spinal cord injury model was established by Allen＇s method in 30 rats.One week after model building,24 rats were randomly divided into 3 groups：the spinal cord injury group,sham group and BMSCs group.Basso,Beattie Bresnahan locomotor rating scale（ BBB） scale was evaluated before intervention,7 d,14 d,21 d,28 d,35 d,and42 d after intervention.The injured spinal cords were obtained at 42 d for pathology test.Results BBB scale in BMSCs group was higher than spinal cord injury group and sham group from 14 d after the intervention.HE stain also showed smaller liquify area and less vesicles.Immunofluorescence showed the transplanted BMSCs can migrate to the injury area,and can differentiate into neuron,glia cells and neural progenitor cells.Conclusion BMSCs can migrate to the injury area in spinal cord injury,and differentiate into nerve cells to promote spinal cord repairing.