Characterization of a primary hepatocyte culture system for toxicological studies
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 dishes in a modified Chee's medium containing 1 microM 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 II 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.
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The model supports extended cell viability and phenotypic maintenance for up to 30 days, as indicated by sustained albumin expression, stable cytochrome P450 activity, and effective ammonia detoxification pathways in untreated samples.
“One of the principal achievements of this work is the model’s capacity to support extended cell viability and phenotypic maintenance for up to 30 days, as indicated by sustained albumin expression, stable cytochrome P450 (CYP450) activity, and effective ammonia detoxification pathways in untreated samples (Zurlo J, 1996; Bachour-El Azzi, 2022; Huch M, 2015) (Figures 1C,D,G).”
From A 3D bioengineered human liver for the study of acute and chronic drug-induced hepatotoxicity and fibrosis (2026) · cites this paper as Zurlo J, 1996
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“…extended cell viability and phenotypic maintenance for up to 30 days, as indicated by sustained albumin expression, stable cytochrome P450 (CYP450) activity, and effective ammonia detoxification pathways in untreated samples (Zurlo J, 1996; Bachour-El Azzi, 2022; Huch M, 2015) (Figures 1C,D,G).”
Jiang, H., Seo, S.-J., Hoshiba, T., Akaike, T., Cho, C., & Biomed, H. (2008)
- Authors
- Zurlo, J., & Arterburn, L. M.
- Affiliations
- Johns Hopkins University (US)
- Grace (United States) (US)
- Venue
- In Vitro Cellular & Developmental Biology - Animal
- Published
- 1996-04
- Volume
- 32
- Issue
- 4
- Pages
- 211-220
- ISSN
- 1071-2690 (ISSN-L)
- 1543-706X
- OpenAlex
- W2070427988
- Cited by
- 35
- References
- 44
- Citations by year
- 2026: 1 · 2017: 1 · 2016: 1 · 2015: 1 · 2014: 1 · 2013: 3 · 2012: 1
- FWCI
- 3.40
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- In Vitro Cellular & Developmental Biology - Animal (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Environmental Health Sciences (ES03819, P30ES003819)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Keywords
- primary hepatocyte culture, rat hepatocytes, collagenase perfusion, dexamethasone, dimethyl sulfoxide, albumin secretion, cytochrome P450 isozymes, testosterone metabolism, diazepam metabolism, CYP1A1/2, CYP2B1/2, CYP2E1, CYP3A, phase II enzymes, glutathione S-transferase, UDP-glucuronosyltransferase, glutathione, hepatocyte viability, in vitro toxicology
- MeSH terms
- Albumins, Animals, Cell Survival, Cytochrome P-450 Enzyme System, Dexamethasone, Diazepam, Dimethyl Sulfoxide, Glutathione, Glutathione Transferase, Mixed Function Oxygenases, Liver, Male, Plastics, Rats, Inbred F344, Testosterone, Glucuronosyltransferase, Toxicity Tests, Cell Culture Techniques, Cytochrome P-450 CYP2B1, Cytochrome P-450 CYP1A1, Rats, Cytochrome P-450 CYP3A
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