Optimal Temperature of Graft Preservation after ex Vivo Gene Transfer in Lung Isografts

Abstract

The aim of this study was to determine the optimal temperature of graft preservation after ex vivo gene transfer to rat lung isografts. Left lungs were harvested and infused with cationic lipid/LacZ-DNA complex via the pulmonary artery, and the grafts were stored for 4h. The grafts (n7) were allocated into groups I-IV according to the storage temperature:4, 10, 16, and 23, respectively. Forty-eight h after orthotopic transplantation, the arterial blood gas was analyzed and the peak airway pressure (PAP) and the level of LacZ protein production in the grafts were measured by reverse transcription polymerase chain reaction. After reperfusion, the grafts were stained with hematoxylin and eosin. The grafts in groups III and IV showed more deterioration as evidenced by decreased arterial oxygen tension, increased PAP, and predominant infiltration of inflammatory cells compared with groups I and II. The level of LacZ production was significantly lower in group I than in groups II-IV. The optimal temperature of lung graft preservation after ex vivo gene transfer was determined to be 10, balancing considerations of lung injury and efficiency of transgene expression.