An improved primary culture method for hippocampal neurons in fetal rats and MAP2 identification

目的：建立一种简单、高效、高纯度的胎鼠海马神经元的原代培养方法.方法：取胎龄18天的Wistar大鼠,显微镜下分离脑组织,采用Brain Dissociation Kit消化获得单个细胞;分别观察培养后24h、3d、5d海马神经元的基本形态结构;培养7d后采用微管蛋白相关标志物2（MAP2）鉴定培养神经元的纯度.结果：此培养方法获得的海马神经元状态良好,生长旺盛.培养第7天,经鉴定,神经元的纯度高达99.62%以上.结论：该方法操作简单、高效,所得神经元纯度高,结果稳定.

Objective： To establish a simple, effective and high-purity primary culture method for fetal rat hippocampalneurons. Methods： Wistar rats of gestational age 18 days were taken and the brain tissue was separated under themicroscope. Single neuronal cells were obtained by digestion with Brain Dissociation Kit, and then were seeded incell plates to observe the basic morphologic structure after 24h, 3d, and 5d. Immunofluorescence of microtubuleassociated protein 2 was applied to assess cell purity of the culture. Results： The hippocampal neurons obtained inthis culture method are in good condition and grow vigorously. On the 7th day after culture, the purity of neuronswas up to 99.62%. Conclusion： The method is simple and effective for obtaining the high-purity and stableneurons.