title: Maintenance of in vivo induced cytochrome P-450s in hepatocyte monolayers at non freezing temperatures.
abstract: Cytochrome P450s (CYPs) induced in rats by 3-methylcholanthrene (3-MC), phenobarbital (PB) and dexamethasone (Dex) were investigated. The inducers had no effect on hepatocyte yield, viability, attachment or spreading on collagen. 3-MC induced ethoxyresorufin deethylase (EROD). Under normothermic conditions the activity fell in culture. However, it was maintained when cells were preserved at 10°C under a gelatin gel. Upon reactivation the activity mirrored that of freshly isolated cells at 37°C. Induced levels were stable for at least 6h , the time to form a confluent monolayer. The investigation was extended to other CYPs by looking at patterns of testosterone metabolism. Phenobarbital had the greatest influence in terms of the quantity and number of metabolites. Culture at 37°C decreased the peaks dramatically within 24 h. All 7 peaks were maintained in the preservation system.
authors: Cardiff School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, United Kingdom. Electronic address: EvansPJ@Cardiff.ac.uk.
authors: Peter J
authors: PJ
authors: Evans
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D065693:Cytochrome P-450 Enzyme Inducers / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology*; D004790:Enzyme Induction; D005615:Freezing; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology*; D010634:Phenobarbital / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013696:Temperature; D013739:Testosterone / Q000378:metabolism
chemical_list: D065693:Cytochrome P-450 Enzyme Inducers; D013739:Testosterone; D008748:Methylcholanthrene; D003907:Dexamethasone; D003094:Collagen; D019363:Cytochrome P-450 CYP1A1; D010634:Phenobarbital
keywords: CYPs; Gelatin; Hypothermic preservation; Preinduced hepatocytes