An improved organ culture method for adult mammalian lung

An improved method for maintaining adult rat lung in submerged organ culture is described in which the alveoli were inflated with agar and 200-micron-thick hand-cut sections were mounted in Rose chambers. The conventional single-compartmented Rose culture chamber was modified by adding a second chamber separated from the first by a gaspermeable membrane. One compartment functioned as an air reservoir and the other housed the explants submerged in nutrient medium. Visking dialysis membrane used underneath the explants prevented cell outgrowth and facilitated the exchange of nutrients and waste products at the glass-tissue interface. Because of the excellent optical properties of the Rose chamber and the thinness of the explants, individual cell types can be identified in the living tissue. The explants were studied with time-lapse cinematography, light microcoscopy, histology, and erythrosine B for dye exclusion. With this modified system the functional life span of the explants was increased from 1 week to 1 month.