贵州医科大学学报

2019, v.44;No.220(01) 59-63

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胰酶稀释法提取乳鼠原代海马神经元
Isolation of Primary Hippocampal Neurons from Newborn Mice Using Diluted Trypsin

康杨婷;王丽琨;伍国锋;
KANG Yangting;WANG Likun;WU Guofeng;Guizhou Medical University;Department of Emergency Neurology,the Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

目的:优化SD乳鼠海马神经元的提取方法。方法:选用出生24 h内的SD乳鼠,分离双侧海马组织,分别用胰蛋白酶稀释法和未稀释法消化后获得细胞悬液;培养细胞,在培养第7天采用神经元特异性烯醇化酶(NSE)免疫荧光染色鉴定神经元数量及纯度。结果:胰酶稀释组提取得到的海马神经元结构特征明显,能形成典型的神经细胞网络,90%以上海马神经细胞呈NSE阳性。结论:用稀释胰蛋白酶消化海马组织并配合特定培养条件可获得生长状态良好、纯度高的原代海马神经细胞。
Objective:To optimize the method for isolating primary hippocampal neurons from newborn mice.Methods:In SD rats newborn within 24 hours of birth,bilateral hippocampus tissues were isolated and digested with 0.25% trypsin and undiluted trypsin(0.125%) to obtain two types of cell suspension,respectively.Cells were cultured.On day 7 of culture,cells were immunofluorescently stained for detecting neuron-specific enolase(NSE) to identify the number and purity of neurons.Results:The isolated hippocampal neurons using 0.125% trypsin had obvious structural features and could form a typical neural cell network.Moreover,over 90% of hippocampal neurons were positive for NSE.Conclusion:Isolated primary hippocampal neurons using 0.125% trypsin had higher purity than using 0.25% trypsin and grew well under proper culture condition.

关键词(KeyWords): 胰酶;稀释技术;神经元,海马;细胞培养;鉴定
trypsin;dilution;neurons,hippocampal;cell culture;identification

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81760245/H0913,81560222/H0913);; 贵州省优秀教育科技人才省长专项资金(1065-09);; 贵州省高层次人才科研特助经费(TZJF-2010-054)

作者(Author): 康杨婷;王丽琨;伍国锋;
KANG Yangting;WANG Likun;WU Guofeng;Guizhou Medical University;Department of Emergency Neurology,the Affiliated Hospital of Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2019.01.011

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