摘要
目的观察大鼠骨髓间充质干细胞(BMMSCs)条件培养基(B-CM))对缺氧大鼠大脑皮质神经元凋亡的影响。方法分离、培养、传代Wistar大鼠BMMSCs,细胞融合达90%时更换培养基,继续培养24h后收集培养上清液即B-CM;取培养8d的新生Wistar大鼠大脑皮质神经元,分正常对照组、缺氧组、B-CM组,分别于缺氧0、6h、复氧24h用四甲基偶氮唑盐微量反应比色(MTT)法检测各组细胞活性,用流式细胞仪和透射电镜技术检测神经元凋亡情况。结果神经元缺氧6h细胞活性较对照组明显降低,复氧24h后细胞凋亡率明显升高(P<0.01)。B-CM培养后细胞活性和缺氧前无差异,细胞凋亡明显较少(P<0.01)。结论B-CM对缺氧-复氧引起的神经元凋亡有明显的防护作用。
Objective To observe the effects of bone marrow mesenchymal stem cells （BMMSCs） conditioned - medium （ B - CM ） on apoptosis in cultured cortical neurons after hypoxia - reoxygenation in vitro in rats. Methods BMMSCs were isolated,cultured and expanded,the culture me-dium was substituted by new medium that contain no serum when cells grew to 90% confluence. After cultured for 24 hours,the medium was collected as B - CM. Cortical neurons of mouse cultured for 8 days were divided into three groups：normal control groups, hypoxic group, B - CM disposed groups. Cell viability were detected by 3 - （ 4,5 - dimethhhiazd - 2 - yl ） - 2,5 - diphenyltetra - zoliu bromidel （MTT） methods and the apoptosis of neurons was examined by flow cytometry and telemicroscopy on hypoxia for 0,6 hours and hypoxia - reoxygenation for 24 hours, respectively. Results Compared with control group, hypoxia for 6 hours could decrease cell viability, hypoxia - reoxygenation for 24 hours increased the cell apoptosis ratio remarkably（P 〈0.01 ） ,and B - CM could ameliorate cell apoptosis（P 〈0.01 ） ,but the cell viability were not affected. Conduslon B - CM can ameliorate neuronal apoptosis induced by bypoxia - reoxygenation.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2007年第14期1068-1069,1071,共3页
Journal of Applied Clinical Pediatrics
基金
甘肃省自然科学基金暨中青年科技基金项目资助(ZSO21-A25-073-Y)
关键词
骨髓间充质干细胞条件培养基
缺氧-复氧
大脑皮质神经元
凋亡
bone marrow mesenchymal stem cells conditoned - medium
hypoxia- reoxygenation
cortical neuron
apoptosis
