Culturing primary hippocampal neurons of neonatal mouse and morphologic observation

VernacularTitle:新生小鼠海马神经元培养及形态学观察的优化方法
Author: Xiang CHANG ; Shuhuan FANG ; Yu ZHANG ; Rong YAN ; Zhao QU ; Xueqin HOU ; Ruyu SU ; Lei ZHANG ; Cong YANG ; Qi WANG
Publication Type:Journal Article
Keywords: hippocampal neurons; calcium phosphate precipitation; GFP transfection; spines
From: Chongqing Medicine 2014;(22):2910-2912
CountryChina
Language:Chinese
Abstract: 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 .