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Titolo:
THE COUMARIN 7-HYDROXYLATION MICROASSAY IN LIVING HEPATIC CELLS IN CULTURE
Autore:
DONATO MT; CASTELL JV; GOMEZLECHON MJ;
Indirizzi:
HOSP UNIV LA FE,CTR INVEST,UNIDAD HEPATOL EXPT,AVDA CAMPANAR 21 VALENCIA 46009 SPAIN HOSP UNIV LA FE,CTR INVEST,UNIDAD HEPATOL EXPT VALENCIA 46009 SPAIN
Titolo Testata:
ATLA. Alternatives to laboratory animals
fascicolo: 2, volume: 26, anno: 1998,
pagine: 213 - 223
SICI:
0261-1929(1998)26:2<213:TC7MIL>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN LIVER-MICROSOMES; O-DEETHYLASE ACTIVITY; RAT HEPATOCYTES; CYTOCHROME-P450-2A ENZYMES; DRUG-METABOLISM; INTACT HUMAN; OXIDATION; PURIFICATION; INHIBITION; TOXICITY;
Keywords:
CYTOCHROME P450 ACTIVITY; COUMARIN 7 HYDROXYLATION; CULTURED HEPATOCYTES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
27
Recensione:
Indirizzi per estratti:
Citazione:
M.T. Donato et al., "THE COUMARIN 7-HYDROXYLATION MICROASSAY IN LIVING HEPATIC CELLS IN CULTURE", ATLA. Alternatives to laboratory animals, 26(2), 1998, pp. 213-223

Abstract

Coumarin 7-hydroxylation was evaluated in hepatic cells from various species, cultured in 96-well plates. This microassay involved incubating living cultured cells with the substrate, followed by fluorimetric quantification of the product released! into the culture supernatant, after hydrolysis of the conjugates of 7-hydroxycoumarin that were formed. Fluorescence was measured directly in the wells by using a microplate fluorescence reader. The major advantages of this technique are its simplicity and automation. the small number of cells required, the reduction in sample handling and assay time, and the possibility of performing repeated assays with the same cell monolayer, since no injury to cells is detectable during the assay. By using this microassay, it was shown that human hepatocytes hydroxylated coumarin at higher ratesthan did rabbit, dog or rat hepatocytes, and that no appreciable metabolic activity was observed in hepatoma cells (Hep G2 and FaO). In addition, methoxsalen was found to be a potent inhibitor of cytochrome P4502A6 activity in living human hepatocytes.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/07/20 alle ore 22:25:24