Long-term maintenance of hepatocytes in primary culture in the presence of DMSO: Further characterization and effect of nafenopin, a peroxisome proliferator

• Published in: Experimental cell research Vol. 171; no. 1; pp. 37 - 51
• Main Authors: Muakkassah-Kelly, S.F., Bieri, F., Waechter, F., Bentley, P., Stäubli, W.
• Format: Journal Article
• Language: English
• Published: Orlando, FL Elsevier Inc 01.07.1987; Elsevier
• Subjects: Animal cells; Animals; Biological and medical sciences; Cell cultures. Hybridization. Fusion; Cells, Cultured; Culture Media; Cytochrome P-450 Enzyme System - biosynthesis; Cytochrome P-450 Enzyme System - metabolism; Dimethyl Sulfoxide - pharmacology; DNA - biosynthesis; Endoplasmic Reticulum - ultrastructure; Enzyme Induction; Fundamental and applied biological sciences. Psychology; Liver - cytology; Liver - drug effects; Liver - metabolism; Liver - ultrastructure; Male; Methylcholanthrene - pharmacology; Microbodies - drug effects; Microbodies - enzymology; Microbodies - ultrastructure; Microscopy, Electron; Molecular and cellular biology; Nafenopin - pharmacology; Oxidoreductases - metabolism; Palmitoyl Coenzyme A - metabolism; Phenobarbital - pharmacology; Propionates - pharmacology; Rats; Rats, Inbred Strains; Cell culture; Vertebrata; Regulation(control); Mammalia; Hepatocyte; Investigation method; Rat; Liver; Rodentia; Peroxisome; Inhibitor; Metabolism
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(87)90249-7

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 β-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 β-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.