Characterization of Precision-cut Rat Lung Slices in a Biphasic Gas/Liquid Exposure System: Effect of O(2)

Influence of oxygen on lung cell differentiation has been studied in precision-cut rat lung slice cultures. Rat lung slices were positioned on rolling inserts placed into vials with opened caps to allow free access to the gaseous phase. This system was placed into a continuous-flow rotating chamber with controlled pO(2), pCO(2) and hygrometry. Slices were cultured in a serum-free medium up to 3 days under an atmosphere of 21 or 70% oxygen. Cellular antioxidant markers demonstrated an oxygen concentration-dependent response. Slices cultured with 70% oxygen exhibited the highest specific activity of catalase, NADPH cytochrome c reductase and gamma-glutamyl transpeptidase (GGT) as well as the highest levels of intracellular glutathione after 48 or 72 hours of incubation. Moreover, hyperoxic exposure altered the expression of lung manganese-containing superoxide dismutase mRNA. Hyperoxia had little or no effect on intracellular ATP levels, which remained high in lung slices compared with freshly isolated tissue. The study of the pulmonary specific functions allowed to confirm maintenance of the in vitro cellular differentiation of lung slices incubated with 21% oxygen: (i) polyamine transport is preserved and exhibited kinetic properties similar to those observed in lung in vivo; (ii) ATP levels remained constant throughout the time course of incubation. This in vitro model proves to be a useful tool to study mechanisms involved after oxygen exposure and will probably be useful for the study of other environmental gaseous contaminants. Further developments in this method are under development.