Abstract
Astrocytes perform critical functions necessary for neuronal survival. Thus, examining the influence of astrocyte function on neuronal cell death during disease, including hypoxia/ischemia, has become an important avenue of investigation. In this chapter we detail the methodology and potential pitfalls for establishing cocultures of astrocytes and cortical neurons for studying hypoxia-induced neuronal death. In brief, astrocyte cultures are first established until they reach confluence. The medium is exchanged from a medium that supports astrocyte growth to a medium that supports neuronal viability 24 h before adding neurons to the astrocyte monolayer. After the neurons mature, the cultures are exposed to severe hypoxia and neuronal death is quantified 1–2 days later.
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Acknowledgments
Part of the work discussed in this chapter was supported in part by: 5K08NS046633 (DAR), 1R01NS054192 (DAR), and 1P01NS050315.
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Rita, G., Emily, T., Rempe, D.A. (2012). Cocultures of Neurons and Astrocytes as a Model for Examining Hypoxia-Induced Neuronal Death. In: Milner, R. (eds) Astrocytes. Methods in Molecular Biology, vol 814. Humana Press. https://doi.org/10.1007/978-1-61779-452-0_23
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DOI: https://doi.org/10.1007/978-1-61779-452-0_23
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