Metabolic and structural viability of precision-cut rat lung slices in culture.
1. The principal objective was to evaluate the functional and structural integrity of precision-cut rat lung slices in culture over 72 h. 2. Lung slices metabolized 7-ethoxycoumarin in a time-dependent fashion, the major metabolites being the sulphate and glucuronide of 7-hydroxycoumarin with very low levels of the free compound. Prior treatment of rats with beta-naphthoflavone elevated markedly the rate of metabolism. The optimum slice thickness, as exemplified by the metabolism of 7-ethoxycoumarin, was about 600 microm. 3. Lung slices retained metabolic viability towards 7-ethoxycoumarin for 8 h, but after this point a marked decline in metabolic activity was noted. However, very low levels of activity were still evident following a 72 h incubation. 4. Morphological examination of lung slices revealed nuclear degeneration and loss of tissue architecture following 24h incubation. When cellular integrity was assessed using lactate dehydrogenase, a time-dependent leakage was evident with maximum loss occurring within 24h; longer incubations did not result in further leakage. 5. It is concluded that precision-cut rat lung slices, of 600 microm thickness, can be maintained metabolically viable in culture for some 8 h.
Umachandran
M
M
Molecular Toxicology Group, School of Biomedical and Molecular Sciences, University of Surrey, Guildford GU2 7XH, UK.
Howarth
J
J
Ioannides
C
C
D000818:Animals; D003374:Coumarins / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008168:Lung / Q000033:anatomy & histology* / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008867:Microtomy; D051381:Rats; D017208:Rats, Wistar; D046509:Tissue Culture Techniques; D019324:beta-Naphthoflavone / Q000494:pharmacology
D003374:Coumarins; C017299:7-ethoxycoumarin; D019324:beta-Naphthoflavone; D007770:L-Lactate Dehydrogenase