接种密度对大鼠海马神经元产出量的影响  

Effects of Plating Density on Rat Hippocampal Neurons Output

作　　者：李一鹏[1,2] 杨凯 刘英富[4] 陈旭义[2,5,4] 刚琳 卢磊[1,2] 陈小义[6] 涂悦[2,5,4] 

机构地区：[1]天津中医药大学,天津300193 [2]天津市神经创伤修复重点实验室,天津300162 [3]锦州医科大学,辽宁锦州121001 [4]武警后勤学院附属医院,天津300162 [5]武警后勤学院附属医院脑创伤与神经疾病研究所,天津300162 [6]武警后勤学院,天津300309

出　　处：《生物技术通讯》2016年第6期838-841,883,共5页Letters in Biotechnology

基　　金：国家自然科学基金青年项目(11102235);天津市科技支撑计划重点项目(14ZCZDGX00500);天津市卫生局科技基金(2013KZ134;2013KZ135);武警后勤学院附属医院种子基金(FYM201432;FYM201417;FYM201542;WHJ2015018);武警后勤学院博士启动基金(WHB201417;WHB201305)

摘　　要：目的：观察不同接种密度对海马神经元细胞数量、形态和神经网络连接的影响,以获得成熟的海马神经元。方法：利用机械分离和胰酶消化的方法获得海马神经元,将所得神经元细胞接种于1×104/cm2、2×104/cm2、4×104/cm2、8×104/cm2,16×104/cm2等5个不同的密度组中,在倒置显微镜镜下观察神经元细胞生长状态,培养5~7 d时行微管相关蛋白2免疫荧光染色鉴定。结果：海马神经元细胞培养7 d后,倒置显微镜下可见在4×104/cm2、8×104/cm2的接种密度下成熟神经元的数量相较其他组差异明显,并且神经元形态典型,建立了丰富的网络连接。结论：适宜的接种密度能提高神经元的成熟数量,为神经元网络的形成和进一步的实验奠定了基础。Objective： To investigate the effects of different inoculation density on hippocampal neurons in cellnumber, morphology and neural network connections to find optimum density to obtain mature hippocampal neu-rons. Methods： Mechanical separation and trypsin digestion method for obtaining hippocampal neurons, resulting inneuronal cells were seeded 1×104/cm2, 2×104/cm2, 4×104/cm2, 8×104/cm2, 16×104/cm2, 5 different density groups. In-verted microscope observed hippocampal neurons growing state. 5~7 days in culture line microtubule-associated pro-tein 2 immunofluorescence staining technique forensic analysis. Results： Hippocampal neurons cultured for 7 daysunder an inverted microscope seeding density observed in 4×104/cm2, 8×104/cm2 by the number of mature neuronssignificant difference compared to other groups, and the typical morphology of neurons, establishing a rich networkconnection. Conclusion： Suitable inoculation density can increase the number of mature neurons, to lay the founda-tion for the formation and further experimental neural network.

关 键 词：细胞培养 海马神经元 接种密度 产出量 

分 类 号：Q813.1[生物学—生物工程]