title: A rat primary hepatocyte culture model for aging studies.
abstract: The purpose of this protocol is to establish a primary hepatocyte culture system as a suitable model to examine age-related changes in Phase II detoxication gene expression. Hepatocytes are isolated using a two-step collagenase perfusion technique from young (3 to 6 months) and old (24 to 28 months) rats and placed in primary culture using collagen (Type I)-coated plates as the extracellular matrix. A supplemented William's E Medium is used as the medium. This culture system maintains hepatocyte viability from both young and old rats for ∼60 hr, as measured by lactate dehydrogenase activity, while also maintaining their respective phenotypes relative to Phase II detoxification. We thus conclude that a collagen-based cell culture system is suitable to study age-associated deficits in Nrf2/ARE-mediated Phase II gene regulation provided that experiments can be conducted within 60 hr after cell isolation.
authors.affiliation: Molecular and Cellular Biology Program, Oregon State University, Corvallis, Oregon, USA.
authors.forename: Swapna V
authors.initials: SV
authors.lastname: Shenvi
authors.forename: Brian M
authors.initials: BM
authors.lastname: Dixon
authors.forename: Kate
authors.initials: K
authors.lastname: Petersen Shay
authors.forename: Tory M
authors.initials: TM
authors.lastname: Hagen
mesh_terms: D000375:Aging; D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000166:cytology* / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D051381:Rats