title: Long-term maintenance of hepatocytes in primary culture in the presence of DMSO: further characterization and effect of nafenopin, a peroxisome proliferator.
abstract: The addition of 2% dimethyl sulfoxide to adult rat hepatocytes cultured in a chemically defined medium at Day 1 after cell plating resulted in maintenance of the cytochrome P-450 content and the cyanide-insensitive palmitoyl-CoA beta-oxidation activity at 66 and 70% of the initial Day 1 values. The addition of phenobarbital, 3-methylcholanthrene, or nafenopin from Day 3 to Day 6 increased the contents of cytochrome P-450 to 128, 239, and 251%, respectively, compared to untreated controls at Day 3. In addition, nafenopin also caused a pronounced and time-dependent increase in palmitoyl-CoA beta-oxidation activity but was found to have only a weak stimulating effect on replicative DNA synthesis (2-fold) when compared to that of epidermal growth factor (6.5-fold). In the presence of dimethyl sulfoxide the hepatocyte cultures could be kept alive for more than 1 month. Exposure of such cultures to nafenopin from Day 1 do Day 37 resulted in survival which was even better than that of their untreated counterparts. This effect was accompanied by the appearance of abundant endoplasmic reticulum membranes and an increased number of peroxisomes.
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D004790:Enzyme Induction; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008830:Microbodies / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D008854:Microscopy, Electron; D009255:Nafenopin / Q000494:pharmacology*; D010088:Oxidoreductases / Q000378:metabolism; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D011422:Propionates / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D003470:Culture Media; D011422:Propionates; D009255:Nafenopin; D010171:Palmitoyl Coenzyme A; D008748:Methylcholanthrene; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C028427:palmitoyl CoA oxidase; D004121:Dimethyl Sulfoxide; D010634:Phenobarbital