Characterization of a primary hepatocyte culture system for toxicological studies

Joanne Zurlo, Linda M. Arterburn

Research output: Contribution to journalArticlepeer-review

Abstract

An hepatocyte culture system was developed for potential use in toxicological studies in vitro. Rat hepatocytes were isolated by two-step collagenase perfusion and cultured on Vitrogen-coated Permanox(TM) dishes in a modified Chee's medium containing 1 μM dexamethasone and 1% dimethylsulfoxide. The cells remained highly viable for at least 10 d as determined by lactate dehydrogenase release and total protein levels. Albumin secretion into the medium, as a measure of differentiated function, was maintained at elevated levels over the course of 10 d in culture. A number of CYP activities were determined by the analysis of testosterone metabolism in freeze-thawed cells, diazepam metabolism in live cells, and specific assays for CYP 1A1/2, 2B1/2, 2E1, and 3A. Results of these assays indicated that a wide range of CYP isozymes were maintained, some activities were enhanced under the conditions of culture and some activities were inducible. Activities of the phase 11 enzymes, glutathione S-transferase and UDP- glucuronosyltransferase, and glutathione levels were also maintained in the cultured hepatocytes for at least 6 d. These results strongly support the use of this hepatocyte culture system for in vitro toxicological studies.

Original languageEnglish (US)
Pages (from-to)211-220
Number of pages10
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume32
Issue number4
DOIs
StatePublished - Apr 1996

Keywords

  • cytochrome P450
  • in vitro toxicology
  • metabolism
  • rat hepatocytes

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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