DONOR LUNG PRESERVATION - EFFECT OF COLD PRESERVATION FLUIDS ON CULTURED PULMONARY ENDOTHELIAL-CELLS

Pulmonary arterial (PA) endothelial cell morphology changes during col d preservation. In the present study, the efficacy of University of Wi sconsin solution (UW), UW solution without colloid (modified UW), Euro -Collins (EC), Marshall's solution (MS), and medium 199 + 10% fetal ca lf serum [culture medium (CM)] in maintaining and regaining the cytosk eleton of cultured porcine PA endothelial cells kept at 4 degrees C an d then rewarmed was compared. Features studied were actin, microtubule s, vinculin, and talin, using immunofluorescence, sodium dodecyl sulfa te-polyacrylamide gel electrophoresis, and immunoblotting; permeabilit y of the cell sheet; wound healing; and phagocytic capacity. When cool ed, microtubules depolymerized in all fluids but EC and MS reduced dep olymerization of actin. Permeability decreased at 4 degrees C (P < 0.0 5), and wound healing and phagocytosis ceased. When rewarmed after EC, UW, and CM preservation, wound healing and phagocytosis started withi n 15 min and proceeded normally. Permeability returned to normal but w as excessive following UW preservation. Microtubule repolymerization w as fastest following UW preservation, and actin filament repolymerizat ion was fastest after EC preservation. Thus the type of preservation f luid used influenced the rate of loss and recovery of specific cytoske letal components, with EC giving the fastest structural and functional recovery.