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.
出处
《生物技术通讯》
CAS
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)
关键词
细胞培养
海马神经元
接种密度
产出量
cell culture
hippocampal neurons
inoculation density
output capacity
