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J Appl Physiol 89: 1553-1560, 2000;
8750-7587/00 $5.00
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Vol. 89, Issue 4, 1553-1560, October 2000

A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation

Jonathan A. Cardella1, Shaf Keshavjee1, Eric Mourgeon1, Stephen D. Cassivi1, Stefan Fischer1, Noritaka Isowa1, Arthur Slutsky2,3, and Mingyao Liu2,3

2 Division of Respiratory Medicine, Departments of 1 Surgery and 3 Medicine, Thoracic Surgery Research Laboratory, Toronto General Hospital, University of Toronto, Toronto, Ontario, Canada M5G 2C4

Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100% O2 at 4°C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37°C) DMEM plus 10% fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia-reperfusion injury at the cellular and molecular level.

organ preservation; cell viability; epithelial cells; acute lung injury


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