UNIT 14.7 A Rat Primary Hepatocyte Culture Model for Aging Studies
Published Online: 1 AUG 2008
Copyright © 2008 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Toxicology
How to Cite
Shenvi, S. V., Dixon, B. M., Petersen Shay, K. and Hagen, T. M. 2008. A Rat Primary Hepatocyte Culture Model for Aging Studies. Current Protocols in Toxicology. 37:14.7:14.7.1–14.7.10.
- Published Online: 1 AUG 2008
- Published Print: AUG 2008
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. Curr. Protoc. Toxicol. 37:14.7.1-14.7.10. © 2008 by John Wiley & Sons, Inc.
- collagen Type 1;
- William's Medium E;
- primary hepatocyte cell culture;
- Phase II detoxification