Inflammation and Reactive Oxygen/Nitrogen Species in Glial/Neuronal Cultures
Inflammation contributes to a wide variety of brain pathologies. In this chapter methods are described for using microglia and astrocytes in culture to investigate inflammatory processes and inflammatory neurodegeneration, including the use of neuronal/glial co-cultures and transwells. Methods for detecting and characterising inflammatory activation in culture include changes in microglial phenotype, microglial phagocytosis and expression of pro-inflammatory cytokines. The protocols for how to determine cellular production of superoxide, hydrogen peroxide, peroxynitrite and nitric oxide in culture are described.
Key wordsInflammation Microglia Pro-inflammatory cytokines Cell specific immunocytochemistry Nitric oxide Peroxynitrite Reactive nitrogen species Reactive oxygen species
Relevant research in our laboratory has been funded by the Wellcome Trust, Medical Research Council, British Heart Foundation, Alzheimer’s Research Trust (UK) and European Commission Joint Research Centre (Italy).
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