2种原代培养方法对新生大鼠大脑皮质神经元生物学特性的影响  

Effects of Two Primary-Cultured Methods on the Biological Characteristics of Cerebral Cortical Neurons from Newly-Born SD Rats

作　　者：王静欢 张志敏 邱炳勋 万东[2] 祝慧凤[1] 

机构地区：[1]西南大学药学院暨中医药学院,重庆400715 [2]重庆医科大学附属第一医院急诊医学科&重症医学科,重庆400016

出　　处：《西南大学学报（自然科学版）》2018年第2期19-26,共8页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(81073084);重庆市自然科学基金项目(CSTC2010BB5127;CSTC2014jcyjA10083);教育部中央高校重点项目(XDJK2012B010)

摘　　要：为明确不同培养方法对新生大鼠大脑皮质神经元成熟时间、形态特征、纯度及活力等生物学特性的影响,采用DMEM培养基加Neurobasal无血清培养基法或Neurobasal无血清培养基法原代培养新生24h内SD大鼠大脑皮质神经元,倒置显微镜下观察细胞形态,MTT法检测细胞活力,免疫荧光细胞化学染色法检测神经元纯度及活性.结果显示,2种方法培养的神经元形态差异无统计学意义;但与DMEM培养基加Neurobasal无血清培养基法相比,Neurobasal无血清培养基法培养的神经元成熟更早,数目更多,纯度更高,活力更强(p<0.05).结果提示,原代培养的大脑皮质神经元部分生物学特性受培养方法直接影响;Neurobasal无血清培养基法所得神经元纯度与活性较高,这为实验目的导向的神经元原代培养方法选择提供了借鉴.To clarify the effects of two primary culture methods on the maturation time span,morphological feature,purity,viability and activity of cerebral cortical neurons,cortical neurons were obtained from newly born SD rats and plated in DMEM ＋ neurobasal and neurobasal nutrient media,respectively.The neuronal morphology was visualized using an inverted contrast microscope,MTT assay was used to evaluate the cellular viability,and immunofluorescence staining was employed to identify the purity and activity of the neurons.No obvious difference between the two methods was observed in their effect on neuronal morphology.However,cellular viability,purity and activity of the neurons in the neurobasal nutrient medium were significantly higher than those of the other method（p〈0.05）.Moreover,the number of cells was greater and the maturation time span was shorter in the neurobasal nutrient medium.These results showed that the biological characteristics of the cerebral cortical neurons in primary culture were directly affected by the culture method.The neurobasal serum-free culture program had higher purity and activity of neurons,which provided a reference for the selection of primary culture methods for neuronal priming.

关 键 词：大脑皮质神经元 原代培养 培养基 纯度 活性鉴定 

分 类 号：R338[医药卫生—人体生理学]