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不同培养方式对大鼠原代神经干细胞自噬的影响

Journal of Apoplexy and Nervous Diseases(2017)

Cited 1|Views9
Abstract
目的 观察比较不同培养方式下神经干细胞(neural stem cells,NSCs)的自噬发生情况.方法 取出生24 h内的Sprague-Dawley(SD)大鼠海马与纹状体,并用细胞球悬浮培养和单层贴壁培养两种方法进行体外培养,在光学显微镜下观察其生长.免疫荧光标记神经干细胞特异性抗原Nestin的表达.透射电镜下观察神经干细胞自噬小体.Western blotting检测自噬相关蛋白LC3Ⅱ、Beclin-1的蛋白表达水平.结果 Nestin在悬浮培养中的阳性率为95%以上,在贴壁培养中接近100%.与贴壁培养比较,悬浮培养自噬体形成增多,自噬标记物LC3Ⅱ/LC3Ⅰ、Beclin-1蛋白表达水平显著提高(差异具有统计学意义P<0.05).结论 成功用悬浮培养和贴壁培养两种方法培养原代神经干细胞;悬浮培养可造成细胞内自噬水平增加,推测其可能的原因为神经球内细胞营养缺乏而激活自噬.
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Key words
Neural stem cells,Autophagy,LC3Ⅱ,Beclin-Rat,Adherent culture
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