啮齿类海马神经元的原代培养及方法改进  

Culturing and Identification of the Neonatal Rodent Hippocampal Neurons

作　　者：陈学周[1] 毕意辉 张可可[1] 张明凯[1] 尤涛[1,2] 

机构地区：[1]安徽医科大学第一附属医院骨科,安徽合肥230022 [2]武汉大学人民医院,湖北武汉430060

出　　处：《安徽农学通报》2016年第2期6-7,13,共3页Anhui Agricultural Science Bulletin

基　　金：国家自然科学基金资助项目(81171797);湖北省自然科学基金资助项目(2011CHB022);中国博士后科学基金(9021346201);安徽省博士后科学基金(9021346202)

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。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 hippoeampal neurons grew well and reaehed a high purity.Conclusion ： Us- ing low concentration of trypsin, differantial adherence and serum-free medium conforms to the requirements of neo- natal rat hippoeampal neurons culture in vitro , so as to provide targeted cells for further research

关 键 词：海马神经元 低浓度胰酶 差速贴壁 无血清培养 

分 类 号：Q42[生物学—神经生物学]