摘要
目的简便、高效地分离和提纯小鼠小脑颗粒神经元,提高培养该神经元的纯度和活性。方法通过采用简便、可行的细胞分离方法,在出生7d的小鼠小脑组织中分离原代神经元。计数分离的原代神经元并分析其活性,免疫荧光染色证实其纯度,观察其胞体和轴突生长情况。结果分离的原代细胞活性可高达95%以上;免疫荧光染色证实神经元占分离细胞的97%;培养24h后的神经元易于观察细胞突起的变化,培养48h后的神经元易于观察轴突的变化。结论该方法方便、可行、高效,可用以提纯小脑颗粒神经元进行体外神经元的生长发育、轴突再生和神经系统疾病的研究。
Objective To find and test an assay that can be used to get cerebellar granule neurons from mouse with high viability and high purity.Methods Cerebellar granule neurons were separated from 7-day old mouse.The cell viability was counted by dyeing the cells with trypan blue,the cell purity was calculated by immunofluorescence staining with βⅢ-tubulin and GFAP,and the morphology of these cells were observed under microscope.Results By using the methods described in this study,we separated cerebellar granule neurons from 7day old mouse,the viability of the cerebellar granule neurons was as high as 95%;double staining with βⅢ-tubulin and GFPA showed that the purity of cerebellar granule neurons could reach 97%.Under microscope,after 24 or 48 hours of in vitro culture,neurite extention or the axons of the cerebellar granule neurons were easy to recognize.Conclusion The cerebellar granule neurons with high viability and high purity extracted by this method can be used for the studies of neuron outgrowth,neurogenesis or other neurological disease in vitro.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2010年第8期621-624,共4页
Journal of China Medical University
基金
辽宁省科技厅社发基金资助项目(2009225010-12)
关键词
原代神经元培养
小脑颗粒神经元
primary neurons culture
cerebellar granule neurons
