【摘要】 目的:研究静脉移植人骨髓间质干细胞对肌萎缩侧索硬化症(ALS)模型小鼠生存期和病理变化的影响。方法:体外培养扩增人骨髓间质干细胞(hMSCs),流式细胞仪鉴定hMSCs的性质及纯度,微量尾静脉血提取模型小鼠DNA,PCR扩增鉴定肌萎缩侧索硬化症模型小鼠(SOD1-G93A阳性小鼠)。将3×106个第5代hM-SCs尾静脉移植入预放疗8周的SOD1-G93A阳性小鼠,用Weyd4分法进行评定移植小鼠和未治疗小鼠的生存期、发病时间,尼氏染色计数脊髓前角运动神经元,组织DNA提取、PCR检测人特异性基因β-globin基因来验证hMSCs在受体小鼠中的植入。结果:生存分析显示尾静脉移植hMSCs的ALS模型小鼠生存期比未治疗小鼠延长18d,延缓发病14d;尼氏染色显示在16周、20周移植小鼠脊髓前角大运动神经元计数多于未治疗小鼠;终末期hMSCs移植小鼠中,在中枢神经系统可检测到人特异性该基因。结论:hMSCs可经过尾静脉移植在ALS小鼠中长期植入,延长生存期,减少脊髓前角运动神经元的丢失,有一定的治疗作用。更多还原

【Abstract】 AIM:To study intravenous transplantation of human mesenchymal stem cells (hMSCs) on the life span and pathological change of SOD1-G93A amyotrophic lateral sclerosis (ALS) mice. METHODS:hMSCs were cultured and expanded from heparinized bone marrow cells from healthy donors and the purity and features were identified with FCM. hMSCs (3×106) resuspended in 0.3 mL DMEM or 0.3 mL DMEM only were injected into the tail vein of genotyped SOD1-G93A ALS mice. The mice were evaluated for signs of motor deficit with 4-point scoring system according to Weydt and the onset and life span were assessed. The pathological change was observed with Nissl staining and number of motor neuron was counted. RESULTS:The onset symptoms in untreated SOD1-G93A ALS mice appeared at (156.6±3.6) d of age and the average life span was (188.3±3.5) d. hMSCs transplantation delayed the onset of ALS type symptoms about 14 d and prolonged the life span about 18 d compared to the untreated SOD1-G93A littermates. The loss of motor neurons in untreated mice was much faster and severer than that in hMSCs transplanted mice. At 16 th week and 20 th week,motor neurons of untreated mice were significantly fewer than those of transplanted mice. β-globin gene in brain was detected in transplanted ALS mice. CONCLUSION:hMSCs migrate to central nervous system after intravenous transplantation,prolong the life span and delay the onset and motor neuron loss in SOD1-G93A ALS mice.更多还原