title: Assessment of cell viability in primary neuronal cultures.
abstract: This unit contains five protocols for assaying cell viability in vitro using primary neuronal cultures, including a novel method for use with transfected neurons. Three of the assays are based on the principle that cell death cascades alter membrane permeability. The lactate dehydrogenase (LDH) release assay measures the amount of the cytoplasmic enzyme released into the bathing medium, while the trypan blue and propidium iodide assays measure the ability of cells to exclude dye from their cytoplasm. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay measures the mitochondrial activity of viable cells by quantifying the conversion of the tetrazolium salt to its formazan product. Finally, the fifth assay details the measurement of luciferase expression as an indication of neuronal viability within a relatively small population of transfected neurons.
authors.affiliation: Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Mandar A
authors.initials: MA
authors.lastname: Aras
authors.forename: Karen A
authors.initials: KA
authors.lastname: Hartnett
authors.forename: Elias
authors.initials: E
authors.lastname: Aizenman
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004396:Coloring Agents / Q000032:analysis / Q000378:metabolism; D006836:Hydro-Lyases / Q000032:analysis / Q000378:metabolism; D007202:Indicators and Reagents / Q000737:chemistry; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013778:Tetrazolium Salts / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D007202:Indicators and Reagents; D013778:Tetrazolium Salts; D008156:Luciferases; D006836:Hydro-Lyases; C045581:lactate dehydratase