摘要
目的:经改良和优化,建立高纯度BALB/c小鼠大脑皮质神经元培养的方法。方法:采用L-多聚赖氨酸包被细胞培养板,取新生BALB/c小鼠(出生24 h内)大脑皮质组织,经0.25%胰酶消化后吹打成单个细胞,按1×106/孔接种于35 mm的六孔板中,用神经元细胞培养种植液培养6 h后换神经元细胞培养饲养液,培养40 h时加入阿糖胞苷抑制神经胶质细胞的生长,随时观察神经元培养情况。结果:培养5 d的神经元细胞形态最为典型;经免疫荧光方法鉴定,神经元细胞纯度为93%。结论:经方法改良与优化,获得了高纯度的原代培养小鼠大脑皮质神经元细胞。
Objective：To establish a method for cultivation of cerebral cortex neuronal cells of newborn BALB/c mice.Methods：The cortexes from newborn（less than 24 h） BALB/c mice were obtained and digested by 0.25% trypsin,and then dissociated into single cell suspension.About 1×10^6 cells were seeded onto each 35 mm dish which was coated by poly-L-lysine overnight previously.After cultivated in seeding medium for 6 h,the neuron cells were cultured in neurobasal medium containing B27,FBS,and glutamine.Cytosine arabinofuranoside was added to the cultures at a final concentration of 5 mg/mL on 40 h.Results：The neuron cells showed a typical morphorlogy at day 5.As indicated by indirect immunofluorescence using antibodies against neuron specific βⅢ tubulin,the purity of the neuronal cultures was 93%.Conlusion：The optimized method to culture neuron from BALB/c mice was established
出处
《生物技术通讯》
CAS
2011年第1期85-88,共4页
Letters in Biotechnology
基金
国家自然科学基金(30771988)
广东省医学科研基金(A2010392)
汕头市科技计划项目(汕府科[2009]70号)
