星形胶质细胞
神经元
生物
海马结构
细胞生物学
分泌物
体外
细胞培养
转基因小鼠
生物神经网络
胚胎干细胞
转基因
神经科学
中枢神经系统
内分泌学
遗传学
基因
作者
Emma V. Jones,Denise Cook,Keith K. Murai
出处
期刊:Methods in molecular biology
日期:2011-11-11
卷期号:: 341-352
被引量:48
标识
DOI:10.1007/978-1-61779-452-0_22
摘要
Astrocytes secrete factors that promote neuron survival, synapse formation, and plasticity. Understanding how these factors perform these roles requires a robust in vitro system that can effectively assess the impact of individual glial factors on neuronal properties. A classical approach to studying neuron-glial interactions in vitro uses a system where dissociated embryonic rat neurons are suspended over a feeder layer of rat astrocytes. Here, we describe a useful "sandwich" co-culture system where postnatal mouse hippocampal neurons are grown suspended above a feeder layer of mouse hippocampal astrocytes. We demonstrate that neurons in these cultures remain healthy beyond 3 weeks in vitro and develop more synapses compared to neurons grown without astrocytes. An advantage of this method is that astrocytes and neurons can be prepared separately from postnatal transgenic or knock-out mouse lines allowing one to study, for example, how wild-type neurons develop in the presence of astrocytes from a knock-out mouse line that lacks the expression of a specific astrocyte-secreted factor. We find this culture system to be a convenient and powerful approach to study the contribution of astrocyte-secreted molecules to neuron development.
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