A Neuron-Astrocyte Co-Culture System to Investigate Astrocyte-Secreted Factors in Mouse Neuronal Development
A Neuron-Astrocyte Co-Culture System to Investigate Astrocyte-Secreted Factors in Mouse Neuronal Development
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DOI：
10.1007/978-1-61779-452-0_22
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发表时间：
2012-01-01
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影响因子：
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通讯作者：
Murai, Keith K.
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文献类型：
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作者：
Jones, Emma V.;Cook, Denise;Murai, Keith K.
星形胶质细胞分泌促进神经元存活、突触形成和可塑性的因子。要了解这些因素如何发挥这些作用，需要一个强大的体外系统，能够有效地评估单个胶质因子对神经元特性的影响。一种在体外研究神经元-神经胶质相互作用的经典方法使用了一种系统，即将分离的胚胎大鼠神经元悬浮在大鼠星形胶质细胞的饲养层上。在这里，我们描述了一种有用的“三明治”共培养系统，在该系统中，出生后的小鼠海马神经元悬浮在小鼠海马星形胶质细胞饲养层的上方生长。我们证明，与没有星形胶质细胞生长的神经元相比，这些培养的神经元在体外培养3周后仍保持健康，并发展出更多的突触。这种方法的一个优点是，可以从出生后的转基因或敲除小鼠品系中分离制备星形胶质细胞和神经元，从而可以研究，例如，在缺乏特定星形胶质细胞分泌因子表达的敲除小鼠品系中，野生型神经元是如何在星形胶质细胞存在的情况下发育的。我们发现这种培养系统是研究星形胶质细胞分泌分子对神经元发育的贡献的一种方便而有力的方法。
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.