Title
A sandwich-cultured rat hepatocyte system with increased metabolic competence evaluated by gene expression profiling
Author
Kienhuis, A.S.
Wortelboer, H.M.
Maas, W.J.
van Herwijnen, M.
Kleinjans, J.C.S.
van Delft, J.H.M.
Stierum, R.H.
TNO Kwaliteit van Leven
Publication year
2007
Abstract
A rapid decline of cytochrome P450 (CYP450) enzyme activities remains a drawback of rat hepatocyte-based in vitro cultures. Consequently, judgment of the toxic potential of compounds that need bioactivation by CYP450s may not be adequate using this model. In the present study, an improved hepatocyte-based in vitro system was developed with special focus on metabolic competence. Therefore, a mixture of CYP450 inducers, phenobarbital, dexamethasone and β-naphthoflavone, was added to culture medium of sandwich-cultured rat hepatocytes. The resulting modified model was evaluated by comparing its genome-wide expression profiles with liver and a standard model without the inducer mixture. Metabolic capacity for CYP450 enzymes showed that the modified model resembled more closely the in vivo situation. Gene expression results revealed large differences between in vivo and both in vitro models. The slight differences between the two sandwich models were predominantly represented by gene expression changes in CYP450s. Importantly, in the modified model, expression ratios of the phase I and the majority of phase II genes more closely resembled liver in vivo. The CYP450 enzyme activities corresponded with gene expression data. In conclusion, for toxicological applications using sandwich-cultured hepatocytes, the modified model may be preferred. © 2007 Elsevier Ltd. All rights reserved.
Subject
Biology
Biomedical Research
Gene expression profiling
In vitro models
Metabolic competence
Primary rat hepatocytes
Sandwich-culture
beta naphthoflavone
cytochrome P450
dexamethasone
phenobarbital
animal cell
article
cell culture
culture medium
enzyme activity
gene expression
gene expression profiling
in vitro study
in vivo study
liver
liver cell
male
metabolic capacity
model
nonhuman
nucleotide sequence
rat
Actins
Animals
Biotransformation
Cytochrome P-450 Enzyme System
Cytological Techniques
Data Interpretation, Statistical
Gene Expression Profiling
Hepatocytes
Hydroxylation
Male
Nucleic Acid Hybridization
Oligonucleotide Array Sequence Analysis
Rats
Rats, Wistar
Reverse Transcriptase Polymerase Chain Reaction
RNA
Testosterone
Rattus
To reference this document use:
http://resolver.tudelft.nl/uuid:b9dc3c02-d1bb-48a7-866b-b65afde5ee58
DOI
https://doi.org/10.1016/j.tiv.2007.01.010
TNO identifier
240122
ISSN
0887-2333
Source
Toxicology in Vitro, 21 (5), 892-901
Document type
article