Astrocytes pp 341-352 | Cite as

A Neuron-Astrocyte Co-Culture System to Investigate Astrocyte-Secreted Factors in Mouse Neuronal Development

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 814)

Abstract

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.

Key words

Hippocampal neurons Astrocytes Synapses Co-culture Astrocyte-secreted factors Synaptogenesis Neuron dissection Postnatal Dendritic spines 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Centre for Research in NeuroscienceMontreal General HospitalMontrealCanada

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