[Hepatic tissue enzymes: study in cultures of rat liver epithelial cells. Control and analysis of cells by cytogenetic and mass spectrometric methods. Pharmacological applications].
Extensive studies of parameters conditioning selection and high plating efficiency of epithelial liver cells at primary seeding allowed us to set up a technique for the routine culture of liver cells from rats of various ages (18 day-old pc to 7 month-old) in Ham F10 medium supplemented with 10 p. cent fetal calf serum and 10 p. cent human serum. Cultures, after several passages, or sometimes at primary seeding were free of fibroblasts. The quality of water for culture medium preparation was found to be a very important parameter. G-banding caryotype showed that cells in culture were diploid until 15-20 passages. Various metabolic pathways have been studied in primary culture and in cell lines: enzymes of the anaerobic metabolism of hexoses and metabolism of steroid hormones and xenobiotics. Activity of glucose-6-phosphatase was nearly lost in all cultures. Activity of glucose-6-phosphatase was nearly lost in all cultures. Aldolase showed a specific liver activity with a cleavage ratio of phosphofructoses (F-1,6-diP/F-1-P) equal to 1 or about 1 in several primary cultures and cell lines. Many metabolites arising from incubation of cell lines with 14C-labelled corticosterone, corticosterone-21-sulfate, testosterone and progesterone have been isolated and quantitated by gas liquid chromatography (GC) and mass fragmentography coupled to GC, using 14C/12C isotope ratio measurements. These metabolites indicate the presence in cultured cells of 3 alpha/beta-steroid-reductases, 4-ene steroid reductases and hydroxylases at various positions: 2 alpha, 2 beta, 6 alpha, 6 beta, 7 alpha, 17 beta and 16 alpha. These cell lines were able to activate carcinogens through the epoxide-diol pathway and are suitable for drug metabolism study.
D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media; D003582:Cytogenetics; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D013058:Mass Spectrometry; D008722:Methods; D051381:Rats
D003470:Culture Media
Enzymes du tissu hépatique : étude dans des cultures de cellules épithéliales de foie de rat. Contrôle et analyse des lignées par des méthodes cytogénétiques et par des méthodes spectrométriques de masse. Applications pharmacologiques.