Human hepatocyte--a model for toxicological studies. Functional and biochemical characterization
Language English Country Slovakia Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
11156444
Knihovny.cz E-resources
- MeSH
- 7-Alkoxycoumarin O-Dealkylase metabolism MeSH
- Antioxidants toxicity MeSH
- Calcium Channel Blockers toxicity MeSH
- Cell Culture Techniques methods MeSH
- Time Factors MeSH
- Cytochrome P-450 CYP1A2 metabolism MeSH
- Cytochrome P-450 CYP3A MeSH
- Adult MeSH
- Hepatocytes chemistry drug effects physiology MeSH
- Enzyme Inhibitors toxicity MeSH
- Microsomes, Liver drug effects MeSH
- Liver drug effects metabolism MeSH
- Cells, Cultured MeSH
- L-Lactate Dehydrogenase metabolism toxicity MeSH
- Middle Aged MeSH
- Humans MeSH
- Nifedipine toxicity MeSH
- Mixed Function Oxygenases metabolism MeSH
- Rifampin toxicity MeSH
- Silymarin toxicity MeSH
- Cytochrome P-450 Enzyme System metabolism MeSH
- tert-Butylhydroperoxide toxicity MeSH
- Toxicity Tests methods MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 7-Alkoxycoumarin O-Dealkylase MeSH
- Antioxidants MeSH
- Calcium Channel Blockers MeSH
- CYP3A protein, human MeSH Browser
- CYP3A4 protein, human MeSH Browser
- Cytochrome P-450 CYP1A2 MeSH
- Cytochrome P-450 CYP3A MeSH
- Enzyme Inhibitors MeSH
- L-Lactate Dehydrogenase MeSH
- Nifedipine MeSH
- Mixed Function Oxygenases MeSH
- Rifampin MeSH
- Silymarin MeSH
- Cytochrome P-450 Enzyme System MeSH
- tert-Butylhydroperoxide MeSH
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.
Identification of Human Sulfotransferases Active towards Silymarin Flavonolignans and Taxifolin
