Influence of medium composition and culture conditions on glutathione S-transferase activity in adult rat hepatocytes during culture.
Glutathione S-transferase (GST) activity was measured in adult rat hepatocytes during either pure culture or coculture with another rat liver cell type in various media. Addition of nicotinamide, selenium, or dimethylsulfoxide, deprivation of cyst(e)ine and the use of two complex media were tested. Whatever the conditions used, after a constant decrease during the first 24 h, GST remained active over the whole culture period (1-2 wk). However, various patterns were observed: GST activity either remained relatively stable to approximately 50% of the initial value or showed a moderate or strong increase. The highest values were found in pure hepatocyte cultures maintained in the presence of nicotinamide or dimethylsulfoxide. Similar changes were observed using 1-chloro-2,4-dinitrobenzene or 1,2-dichloro-4-nitrobenzene as substrates for GST. Addition of 10(-4) M indomethacin resulted in 37 to 60% inhibition of enzyme activity. Thus, these results demonstrate that GST remained expressed during culture but its levels markedly varied depending on the medium composition and type and age of culture.
Vandenberghe
Y
Y
INSERM, Unité de Recherches Hépatologiques U 49, Hôpital de Pontchaillou, Rennes, France.
Ratanasavanh
D
D
Glaise
D
D
Guillouzo
A
A
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media; D003545:Cysteine / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007213:Indomethacin / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000201:enzymology*; D009536:Niacinamide / Q000494:pharmacology; D051381:Rats; D012643:Selenium / Q000494:pharmacology
D003470:Culture Media; D009536:Niacinamide; D003577:Cytochrome P-450 Enzyme System; D005982:Glutathione Transferase; D012643:Selenium; D003545:Cysteine; D007213:Indomethacin; D004121:Dimethyl Sulfoxide