脑源性神经营养因子基因修饰脂肪间充质干细胞治疗大鼠急性脊髓损伤
Brain derived neural growth factor gene-modified adipose derived mesenchymal stem cells in treating acute rat spinal cord injury
目的 观察脑源性神经营养因子(BDNF)基因修饰脂肪间充质干细胞(ADSCs)治疗大鼠急性脊髓损伤效果.方法 获取大鼠ADSCs细胞,观察其生物学特征,行溴脱氧尿嘧啶核苷(BrdU)标记及腺病毒BDNF基因转染.将36只SD大鼠随机分为3组:脊髓损伤对照组(A)、ADSCs治疗组(B)及BDNF转染的ADSCs治疗组(C).在脊髓损伤部位行相应处理:A组植入明胶海绵;B组植入含3.0μl(3×1010/L) ADSCs的明胶海绵;C组植入含3.0μl(3×1010/L)转染ADSCs的明胶海绵.行为学评分及观察免疫荧光染色.结果 成功分离并培养ADSCs,可于体外稳定增殖.术后7、14、28 d,C组运动功能评分为(10.55±0.80、14.39 ±0.25、19.71 ±2.14)分,明显高于A组(5.35 ±0.68、7.93±0.45、10.78 ±1.18)分、B组(7.01 ±0.39、10.33±0.27、14.87 ±0.37)分.免疫荧光染色显示移植ADSCs脊髓组织内,均可见不同程度的分布有BrdU阳性标记细胞;荧光染色结果提示部分BrdU阳性的移植细胞同时呈胶质纤维酸性蛋白(GFAP)阳性.结论 ADSCs在移植后能在宿主体内存活、迁移并分化.移植ADSCs之后,运动功能可见改善,证实ADSCs移植能改善中枢神经系统功能.
更多Objective To observe the effect of brain derived neural growth factor (BDNF) genemodified adipose derived mesenchymal stem cells (ADSCs) in treating acute rat spinal cord injury (SCI).Methods Rat ADSCs were harvested and labeled with BrdU.The ADSCs were genetically modified with Adv-BDNF.Thirty-six SD rats were randomly divided into SCI group (A),ADSCs group (B) and BDNFADSCs group (C),subject to implantation of gelatin sponge,gelatin sponge with ADSCs,and gelatin sponge with BDNF-ADSCs,respectively.Behavior score and immunofluorescence staining were observed.Results The ADSCs were cultured and labeled with BrdU successfully.On the postoperative day 7,14,and 28,the blood brain barrier(BBB) scores in group C were 10.55 ±0.8,14.39 ±0.25 and 19.71 ±2.14,which were obviously higher than in group A (5.35 ±0.68,7.93 ±0.45,10.78 ± 1.18) and group B (7.01 ±0.39,10.33 ±0.27,14.87 ±0.37).The immunofluorescence assay showed that the transplanted ADSCs migrated to the injured site and differentiated into neuron-like cells.The neurological function was greatly improved after the ADSCs transplantation.Conclusion The ADSCs can survive,migrate to the injured site in the spine and differentiate into neuron-like cells.The motor function is greatly improved after the transplantation.
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