Functional integrity of precision-cut liver slices from deer and cattle.
Precision-cut bovine and cervine liver slices were incubated in RPMI 1640 media for up to 72 h, and cellular integrity was assessed utilizing the leakage of lactate dehydrogenase (LDH) and the formation of the formazan metabolite of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT). Leakage of LDH (80%) from the bovine and cervine slices was noted only following 10 h of culture, and similarly, the generation of MTT-formazan declined. Metabolic viability was determined using 7-ethoxycoumarin as the model substrate, which was metabolized by slices from both animal species in a time-dependent manner for at least 6 h to generate 7-hydroxycoumarin, which was secreted into the media primarily as glucuronide and sulphate conjugates. With both cervine and bovine slices metabolic activity decreased markedly after a 4-h preincubation as assessed following a further 2-h incubation in the presence of 7-ethoxycoumarin. Subsequently, ethoxycoumarin metabolism by bovine slices did not decrease further until 24 h and was still detectable at 72 h. In the case of cervine slices, activity declined gradually after 8 h, with no activity being detectable at 72 h. It may be concluded that cervine and bovine slices may be maintained metabolically active for 8-10 h, which should prove sufficient for xenobiotic metabolism studies to be performed.
Sivapathasundaram
S
S
Molecular Toxicology Group, School of Biomedical and Life Sciences, University of Surrey, Guildford GU2 7XH, Surrey, UK.
Howells
L C
LC
Sauer
M J
MJ
Ioannides
C
C
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003374:Coumarins / Q000378:metabolism*; D003670:Deer; D005562:Formazans / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D008297:Male; D013045:Species Specificity; D013778:Tetrazolium Salts / Q000378:metabolism*
D003374:Coumarins; D005562:Formazans; D013778:Tetrazolium Salts; C079782:MTT formazan; C017299:7-ethoxycoumarin; D007770:L-Lactate Dehydrogenase