Unit

UNIT 14.7 A Rat Primary Hepatocyte Culture Model for Aging Studies

  1. Swapna V. Shenvi1,2,
  2. Brian M. Dixon1,2,
  3. Kate Petersen Shay2,
  4. Tory M. Hagen2,3

Published Online: 1 AUG 2008

DOI: 10.1002/0471140856.tx1407s37

Current Protocols in Toxicology

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.

Author Information

  1. 1

    Molecular and Cellular Biology Program, Oregon State University, Corvallis, Oregon

  2. 2

    Linus Pauling Institute, Oregon State University, Corvallis, Oregon

  3. 3

    Department of Biochemistry and Biophysics, Oregon State University, Corvallis, Oregon

Publication History

  1. Published Online: 1 AUG 2008
  2. Published Print: AUG 2008

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. Curr. Protoc. Toxicol. 37:14.7.1-14.7.10. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • collagen Type 1;
  • William's Medium E;
  • primary hepatocyte cell culture;
  • aging;
  • Phase II detoxification