Isolated human hepatocytes (HH) are an accepted model for in vitro experiments for testing liver function and xenobiotic metabolism. Preferred over more traditional animal hepatocyte model used in toxicological studies, it is the model of choice when substances undergoing biotransformation in man are investigated. The aim of this study was to optimize isolation and culture conditions for HH primary culture with regard to cell yield, viability, and metabolic activity, and to evaluate the suitability of donor samples for toxicology experiments. Cell viability, total cytochrome P450 (CYP) content, CYP3A4, CYP1A2 activity, and finally mixed ethoxycoumarin-O-deethylase (ECOD) activity were parameters measured in order to characterize the isolated HH. The quality of the primary cultures, stable and functional for a seven-day period following 24 hour stabilization, was assessed by lactate dehydrogenase (LDH) leakage and response to the model toxin tert-butylhydroperoxide (tBH) and to silybinin, a model cytoprotective substance. Based on HH obtained from livers of five multiorgan donors (average age 44.8 years, three males and two females), the individual variability of donors needs to be considered in evaluating cultures focussing on clinical liver tests. Greater sensitivity to toxins and silybinin was found in the hepatocyte culture from one donor with higher aminotransferase activity. In another case, higher serum bilirubin appeared to be linked to higher ECOD activity. Our conclusion is that values of clinical liver tests ought to suggest a healthy organ thus eliminating previous hepatocyte damage, the crucial factor of primary culture stability and functioning.

• MeSH
• 7-alkoxykumarin-O-dealkylasa metabolismus   MeSH
• antioxidancia toxicita   MeSH
• blokátory kalciových kanálů toxicita   MeSH
• buněčné kultury metody   MeSH
• časové faktory MeSH
• cytochrom P-450 CYP1A2 metabolismus   MeSH
• cytochrom P-450 CYP3A MeSH
• dospělí MeSH
• hepatocyty chemie   účinky léků   fyziologie   MeSH
• inhibitory enzymů toxicita   MeSH
• jaterní mikrozomy účinky léků   MeSH
• játra účinky léků   metabolismus   MeSH
• kultivované buňky MeSH
• L-laktátdehydrogenasa metabolismus   toxicita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nifedipin toxicita   MeSH
• oxygenasy se smíšenou funkcí metabolismus   MeSH
• rifampin toxicita   MeSH
• silymarin toxicita   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• terc-butylhydroperoxid toxicita   MeSH
• testy toxicity metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 7-alkoxykumarin-O-dealkylasa MeSH
• antioxidancia MeSH
• blokátory kalciových kanálů MeSH
• CYP3A protein, human MeSH Prohlížeč
• CYP3A4 protein, human MeSH Prohlížeč
• cytochrom P-450 CYP1A2 MeSH
• cytochrom P-450 CYP3A MeSH
• inhibitory enzymů MeSH
• L-laktátdehydrogenasa MeSH
• nifedipin MeSH
• oxygenasy se smíšenou funkcí MeSH
• rifampin MeSH
• silymarin MeSH
• systém (enzymů) cytochromů P-450 MeSH
• terc-butylhydroperoxid MeSH