Liver slices in dynamic organ culture. II. An in vitro cellular technique for the study of integrated drug metabolism using human tissue.
1. Precision cut human liver slices in dynamic organ culture have been used to study the integrated metabolism of 7-ethoxycoumarin and the conjugation of 7-hydroxycoumarin. 2. The metabolism of 7-ethoxycoumarin and 7-hydroxycoumarin was monitored for 6 h. For both substrates there was a time-dependent increase in metabolites present in the incubation medium. The low levels of free 7-hydroxycoumarin found in the medium when 7-ethoxycoumarin was the substrate suggests good coupling of phase I and phase II metabolism. 3. With suitable incubation conditions, i.e. change of medium containing new substrate every 2 h, the metabolism of both 7-ethoxycoumarin and 7-hydroxycoumarin by human liver slices was found to proceed at similar rates for up to 24 h. This was demonstrated using five separate human liver preparations. 4. Human liver slices also metabolized mono-chlorobenzene and o-, m- and p-dichlorobenzene to aqueous soluble metabolites. There was a time-dependent increase in the appearance of aqueous soluble metabolites present in the incubation medium. Metabolites were not retained by the liver slices. 5. A cold-storage transit buffer has been described and used to maintain the levels of drug metabolism in both rat and human tissue for periods of up to 6 h. 6. The use of human liver slices in dynamic organ culture as a suitable method for the direct assessment of integrated hepatic drug metabolism is proposed.
Barr
J
J
Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson 85721.
Weir
A J
AJ
Brendel
K
K
Sipes
I G
IG
D000293:Adolescent; D000328:Adult; D000818:Animals; D001711:Biotransformation; D002648:Child; D002675:Child, Preschool; D002722:Chlorobenzenes / Q000378:metabolism; D003374:Coumarins / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005260:Female; D005965:Glucuronates / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008297:Male; D009924:Organ Culture Techniques; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D013431:Sulfates / Q000378:metabolism; D014468:Umbelliferones / Q000378:metabolism
D002722:Chlorobenzenes; D003374:Coumarins; D005965:Glucuronates; D004364:Pharmaceutical Preparations; D013431:Sulfates; D014468:Umbelliferones; C017299:7-ethoxycoumarin; C031477:7-hydroxycoumarin; C004726:2-dichlorobenzene; C042146:1,3-dichlorobenzene; D003577:Cytochrome P-450 Enzyme System; C018511:4-dichlorobenzene