三种皮质神经元原代培养方法的改良及对比

发布时间：2018-03-26 17:50

本文选题：皮质神经元　切入点：大鼠　出处：《中国实验动物学报》2015年04期

【摘要】：目的改良并比较机械分离法、胰蛋白酶消化法和木瓜蛋白酶消化法在大鼠皮质神经元培养中的优缺点,为科技工作者根据各自条件选择合适的培养方法提供理论依据。方法采用胎龄为16~18 d的SD大鼠胚胎,分别用机械分离、胰酶消化以及木瓜蛋白酶消化三种方法对SD胎鼠皮层神经元进行培养,并对不同培养时间的神经元进行形态学观察和免疫荧光显微镜纯度鉴定。结果三种方法均能成功培养出杂质少且纯度高的皮层神经元,其在不同生长阶段具有典型的形态学特征。经NSE免疫荧光染色,神经元纯度分别在96.28%,95.63%及97.34%,三组间无统计学差异(P0.05)。结论三种方法均成功培养出杂质量较少且纯度高的皮层神经元,均可作为神经元体外培养的良好实验模型,且三种培养方法都有其不同的特点。
[Abstract]:Objective to compare the advantages and disadvantages of mechanical separation, trypsin digestion and papain digestion in rat cortical neuron culture. To provide theoretical basis for scientific and technological workers to select suitable culture methods according to their respective conditions. Methods SD rat embryos with gestational age of 16 ~ 18 days were mechanically separated. The cortical neurons of SD fetal rats were cultured by trypsin digestion and papain digestion. Morphological observation and immunofluorescence microscopy were carried out on the neurons cultured for different time. Results the cortical neurons with low impurity and high purity could be successfully cultured by the three methods. NSE immunofluorescence staining showed that the purity of neurons was 95.63% and 97.34%, respectively, and there was no significant difference among the three groups (P 0.05). Conclusion the cortical neurons with low impurity and high purity were successfully cultured by the three methods. It can be used as a good experimental model of neuron culture in vitro, and all three methods have their own characteristics.
【作者单位】：中南大学湘雅医学院基础医学院法医系;
【基金】：中南大学中央高校基本科研业务费专项资金资助(项目代码:502200372)
【分类号】：R329.2

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