title: Cocultures of neurons and astrocytes as a model for examining hypoxia-induced neuronal death.
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.
authors.affiliation: Center for Neural Development and Disease, University of Rochester School of Medicine and Dentistry, Rochester, NY, USA.
authors.forename: Giuliano
authors.initials: G
authors.lastname: Rita
authors.forename: Terho
authors.initials: T
authors.lastname: Emily
authors.forename: David A
authors.initials: DA
authors.lastname: Rempe
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods*; D016923:Cell Death / Q000502:physiology*; D003470:Culture Media / Q000737:chemistry*; D000860:Hypoxia / Q000473:pathology*; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology* / Q000473:pathology
chemical_list: D003470:Culture Media