Interspecies difference in liver-specific functions and biotransformation of testosterone of primary rat,porcine and human hepatocyte in an organotypical sandwich culture |
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Authors: | Angelika Langsch,Shibashish Giri,Ali Acikgö z,Inka Jasmund,Bernd Frericks,Augustinus Bader |
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Affiliation: | 1. Hanover Medical School, 30625 Hannover, Germany;2. Cell Techniques and Applied Stem Cell Biology, Biomedical-Biotechnological Center (BBZ), Deutscher Platz 5, D-04103 Leipzig, Germany |
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Abstract: | Interspecies difference is an important issue in toxicology research. We compared the potential in vitro metabolism of human, porcine and rat hepatocytes over 2 weeks in culture in an organotypical culture model which reflects the in vivo situation. All three species show similar LDH-rates. Albumin measurements showed that rat cells are about twice as active as human and porcine hepatocytes. The ethoxyresorufin-O-deethylase (EROD) activity of the rat hepatocytes is with about 14 μU/106 cells distinctly higher than those of porcine and human cells (1.8 and 0.5 μU/106 cells respectively), furthermore, the activity of the rat EROD increases slightly during the prolonged time in culture, whereas those of porcine and human enzymes slightly decrease. Concerning ethoxycoumarin-O-deethylase (ECOD), the enzyme activities are found to be in three different ranges where rat cells show the highest activity with 66 μU/106 cells, porcine hepatocytes exhibit an activity of about 23 μU/106 cells, and human activity is lowest with 0.7 μU/106 cells. All three species show a similar decreasing trend of ECOD during the period of study. Regarding the biotransformation of testosterone, human and porcine liver cells form three major metabolites whereas rat cells form a mixture of all measured metabolites. Hence, in vitro metabolism using porcine hepatocytes would be much more scientific sense than one using rat hepatocytes since the metabolic pathways are much closer to human metabolism. |
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Keywords: | BSA, bovine serum albumin CYP, cytochrome P450 DMEM, Dulbecco's Modified Eagle's Medium EGTA, ethylene glycol-bis(β-aminoethyl ether)N,N,N&prime ,N&prime -tetraacetic acid HBSS, Hanks&rsquo balanced salt solution HEPES, 2-[4-(hydroxyethyl)-1-piperazino]-ethansulfonic acid LDH, lactate dehydrogenase WE, William's Medium E EROD, ethoxyresorufin-O-deethylase ECOD, ethoxycoumarin-O-deethylase FCS, fetal calf serum TMB, tetramethylbenzidine FDA, Food and Drug Administration |
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