The use of precision-cut rat lung slices for studying PGF2α metabolism

Abstract The suitability of a dynamic lung slice culture system as an in vitro model for studying pulmonary metabolism of PGF 2 α was assessed. [ 3 H]Prostaglandin F 2 α ([ 3 H]PGF 2 α ), a twenty carbon fatty acid that contains a five-carbon ring and is known to be metabolized by lung in vivo, was incubated with precision-cut rat lung slices in 1.7 ml of Waymouth's buffer fortified with 10% fetal calf serum. At 0, 2, 4 and 8 h after addition of [ 3 H]PGF 2 α (1.82 ng/μCi), incubation was stopped and the contents of each vial were analyzed for [ 3 H]PGF 2 α and its metabolites using reversed-phase HPLC with radiochemical detection. PGF 2 α was metabolized to 15-keto PGF 2 α , 13,14-dihydro-15-keto PGF 2 α , and two unknown minor polar metabolites. These results indicate that PGF 2 α was metabolized in lung slices pathways similar to those seen in vivo. Slice viability was assessed by protein synthesis and light microscopic examination of lung slices through 24 h of incubation. Protein synthesis was maintained and no tissue necrosis was observed over the entire 24 h incubation, indicating that the lung slices were viable for at least 24 h. These results indicate that the dynamic lung slice culture system is an appropriate in vitro model for studying the pulmonary metabolism of PGF 2 α .