Document Detail


Human hepatocyte--a model for toxicological studies. Functional and biochemical characterization.
MedLine Citation:
PMID:  11156444     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
M Modrianský; J Ulrichová; P Bachleda; P Anzenbacher; E Anzenbacherová; D Walterová; V Simánek
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  General physiology and biophysics     Volume:  19     ISSN:  0231-5882     ISO Abbreviation:  Gen. Physiol. Biophys.     Publication Date:  2000 Jun 
Date Detail:
Created Date:  2001-01-11     Completed Date:  2001-03-29     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8400604     Medline TA:  Gen Physiol Biophys     Country:  Slovakia    
Other Details:
Languages:  eng     Pagination:  223-35     Citation Subset:  IM    
Affiliation:
Department of Medical Chemistry and Biochemistry, Medical Faculty, Palacký University, Olomouc, Czech Republic. oregon@tunw.upol.cz
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MeSH Terms
Descriptor/Qualifier:
7-Alkoxycoumarin O-Dealkylase / metabolism
Adult
Antioxidants / toxicity
Calcium Channel Blockers / toxicity
Cell Culture Techniques / methods*
Cells, Cultured
Cytochrome P-450 CYP1A2 / metabolism
Cytochrome P-450 CYP3A
Cytochrome P-450 Enzyme System / metabolism
Enzyme Inhibitors / toxicity
Female
Hepatocytes / chemistry*,  drug effects*,  physiology*
Humans
L-Lactate Dehydrogenase / metabolism,  toxicity
Liver / drug effects,  metabolism
Male
Microsomes, Liver / drug effects
Middle Aged
Mixed Function Oxygenases / metabolism
Nifedipine / toxicity
Rifampin / toxicity
Silymarin / toxicity
Time Factors
Toxicity Tests / methods
tert-Butylhydroperoxide / toxicity
Chemical
Reg. No./Substance:
0/Antioxidants; 0/Calcium Channel Blockers; 0/Enzyme Inhibitors; 0/Silymarin; 13292-46-1/Rifampin; 21829-25-4/Nifedipine; 75-91-2/tert-Butylhydroperoxide; 9035-51-2/Cytochrome P-450 Enzyme System; EC 1.-/Mixed Function Oxygenases; EC 1.1.1.27/L-Lactate Dehydrogenase; EC 1.14.13.-/7-Alkoxycoumarin O-Dealkylase; EC 1.14.13.67/CYP3A4 protein, human; EC 1.14.14.1/CYP3A protein, human; EC 1.14.14.1/Cytochrome P-450 CYP1A2; EC 1.14.14.1/Cytochrome P-450 CYP3A

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