Donor lung preservation: effect of cold preservation fluids on cultured pulmonary endothelial cells

Abstract

Pulmonary arterial (PA) endothelial cell morphology changes during cold preservation. In the present study, the efficacy of University of Wisconsin solution (UW), UW solution without colloid (modified UW), Euro-Collins (EC), Marshall's solution (MS), and medium 199 + 10% fetal calf serum [culture medium (CM)] in maintaining and regaining the cytoskeleton of cultured porcine PA endothelial cells kept at 4 degrees C and then rewarmed was compared. Features studied were actin, microtubules, vinculin, and talin, using immunofluorescence, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting; permeability of the cell sheet; wound healing; and phagocytic capacity. When cooled, microtubules depolymerized in all fluids but EC and MS reduced depolymerization of actin. Permeability decreased at 4 degrees C (P <0.05), and wound healing and phagocytosis ceased. When rewarmed after EC, UW, and CM preservation, wound healing and phagocytosis started within 15 min and proceeded normally. Permeability returned to normal but was excessive following UW preservation. Microtubule repolymerization was fastest following UW preservation, and actin filament repolymerization was fastest after EC preservation. Thus the type of preservation fluid used influenced the rate of loss and recovery of specific cytoskeletal components, with EC giving the fastest structural and functional recovery.