| 啮齿类海马神经元的原代培养及方法改进 |
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| 引用本文： | 陈学周,毕意辉,张可可,张明凯,尤涛.啮齿类海马神经元的原代培养及方法改进[J].安徽农学通报,2016,22(2). |
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| 作者姓名： | 陈学周 毕意辉 张可可 张明凯 尤涛 |
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| 作者单位： | 安徽医科大学第一附属医院骨科,安徽合肥,230022;安徽医科大学第一附属医院骨科,安徽合肥 230022;武汉大学人民医院,湖北武汉 430060 |
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| 基金项目： | 国家自然科学基金资助项目，湖北省自然科学基金资助项目，中国博士后科学基金，安徽省博士后科学基金 |
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| 摘 要： | 目的:通过对新生啮齿类海马神经元的分离和培养,尝试建立一个简单、稳定、高效的啮齿类海马神经元原代培养方法,为脊髓损伤的相关分子机制研究提供目的细胞。方法:取新生SD乳鼠的海马组织,通过低浓度胰酶消化制成细胞悬液、4h差速贴壁后使用无血清Neurobasal培养基培养,倒置显微镜观察细胞生长状态,免疫荧光对海马神经元相关微管蛋白-2(MAP2)行特异性染色,结合DAPI核染色鉴定神经元。结果:该方法培养的海马神经元生长状态良好,纯度较高。结论:采用低浓度胰酶消化,差速贴壁及无血清培养啮齿类海马神经元符合体外细胞实验要求,为进一步研究提供良好的目的细胞。
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| 关 键 词： | 海马神经元 低浓度胰酶 差速贴壁 无血清培养 |
Culturing and Identification of the Neonatal Rodent Hippocampal Neurons |
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| Abstract: | Obejctive:To established a simple,effective and stable method for the culture of rat hippocampal neurons in vitro,and provide target cell for dissecting molecular and cellular mechanisms in neuroscience.Methods:The hip-pocampus of neonatal rat were dissected and cell suspension were digested by low concentration of trypsin.The cell suspensions were cultured with serum-free Neurobasal medium after four hours differential adherence.Morphological observation of neurons growth state by inverted microscope,the hippocampal neurons can be identified by MAP2 specific antibody with DAPI.Results:The hippocampal neurons grew well and reached a high purity.Conclusion:Us-ing low concentration of trypsin,differantial adherence and serum-free medium conforms to the requirements of neo-natal rat hippocampal neurons culture in vitro,so as to provide targeted cells for further research. |
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| Keywords: | Hippocampal neurons Low concentration of trypsin Differantial adherence Serum-free medium |
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