摘要
目的:建立胎鼠、新生鼠海马神经元体外培养方法。方法:分别取胎鼠、新生鼠海马,消化后种植,用含有2%B27的neurobasal培养液培养,第3天加入5μmol·L^(-1)的阿糖胞苷,换液后继续培养,以获得纯度较高的海马神经元。培养第3、7天观察细胞生长及突起情况,用neurofilament抗体以免疫荧光方法鉴定神经元细胞。结果:海马神经元种植24 h后贴壁,7天时神经元突起相互连接成网络。经neurofilament染色,培养细胞阳性率高,新生鼠原代培养海马神经元阳性率达(89±3.4)%,胎鼠海马神经元阳性率达(98±1.5)%。结论:本方法培养出的胎鼠、新生大鼠海马神经元纯度较高,可作为海马神经元模型用于进一步研究。
Objective： To establish the method to culture hippocampal neurons from fetal rats and newborn rats. Methods： The hippocampus of new born and fetal SD rats was dissected and digested with trypsin,then was cultivated in Neurobasal 20% B27 free serum culture system. After 3 days,adding cytarabine to purify neuron. The morphology of hippocampus neuron was observed on l day,3 day and 7 day by the inverted microscope; the purity quotient and growth state of neuron cell was observed on 7 day by immunofluorescence. Results： The hippocampus neurons adhered in 24 hours. Neurites of hippocampus neurons connected in 7 days. Cells were identified by Neurofilament staining. The purity degree of new born rat hippocampus neurons was about（87 ± 3. 4） % and The purity degree of fetal rat was about（98 ± 1. 5） %.Conclusion： Neurobasal 20% B27 free serum culture system with cytarabine is a good method of culture and purify hippocampus neuron cell from fetal rats and newborn rats.
出处
《赣南医学院学报》
2017年第3期354-356,共3页
JOURNAL OF GANNAN MEDICAL UNIVERSITY
关键词
海马神经元
细胞培养
胎鼠
新生鼠
hippocampus neuron
cell culture
fetal rat
newborn rat
