CAJ | 학술논문

目的：探讨一种更加优化的体外培养海马神经元及进行形态学观察的方法，为阿尔茨海默病（A D ）神经突触的研究奠定基础。方法选择新出生后0～1 d的C57BL/6J小鼠，断头取双侧海马，采用低浓度胰酶消化加机械分离的方法，使用无血清培养基进行神经元原代培养，培养17 d后利用磷酸钙共沉淀法进行绿色荧光蛋白（GFP ）的转染，于荧光显微镜下观察海马神经元和树突棘形态特征。结果采用此方法进行的海马神经元原代细胞培养，神经元生长良好，转染GFP后可以看到清晰的轴突/树突和树突棘突等典型的神经细胞结构特征。结论此技术方法所培养的海马神经元生长良好，磷酸钙共沉淀法转染GFP后能够更加清晰地观察到神经元及树突棘的形态特征。
목적：탐토일충경가우화적체외배양해마신경원급진행형태학관찰적방법，위아이자해묵병（A D ）신경돌촉적연구전정기출。방법선택신출생후0～1 d적C57BL/6J소서，단두취쌍측해마，채용저농도이매소화가궤계분리적방법，사용무혈청배양기진행신경원원대배양，배양17 d후이용린산개공침정법진행록색형광단백（GFP ）적전염，우형광현미경하관찰해마신경원화수돌극형태특정。결과채용차방법진행적해마신경원원대세포배양，신경원생장량호，전염GFP후가이간도청석적축돌/수돌화수돌극돌등전형적신경세포결구특정。결론차기술방법소배양적해마신경원생장량호，린산개공침정법전염GFP후능구경가청석지관찰도신경원급수돌극적형태특정。
Objective To discuss a optimal culture method of primary hippocampal neurons and a more suitable method of mor-phological observation ,and provide basis to the study of synapse in Alzheimer′s Disease .Methods Postnatal 0 -1 days (P0 -1 ) C57BL/6J mice were decollated and bilateral hippocampus were separated .Low level concentration of trypsin and mechanical disso-ciation were adopted .And culture medium without serum was used to culture neurons .After 17 days culturing ,transfected neurons with Green Fluorescent Protein(GFP) by calcium phosphate precipitation ,and then observed neurons and spines by fluorescence mi-croscope .Results The neurons looked good and healthy by using this method .And the axons ,dendrites and spines which were typ-ical structure of neurons were observed clearly after transfected with GFP .Conclusion The cultured hippocampal neurons look good by this method .And the morphological characteristics of neurons and spines are observed much more clearly after transfected GFP by calcium phosphate precipitation .