Background: Ex vivo lung perfusion (EVLP) is a promising technique that allows organ preservation and repair, increasing the number of marginal donor lungs for lung transplantation. This study aimed to establish an EVLP model in mice, and to unveil the molecular mechanisms responsible of EVLP beneficial effects. Methods: The mouse EVLP system was established, and mouse lungs were exposed to: (1) without ischemia or EVLP (control), (2) 45 min EVLP followed by 135 min cold ischemia (EVLP-CI), (3) 135 min cold ischemia followed by 45 min EVLP (CI-EVLP), and (4) 180 min cold ischemia (CI). At the end of the 3-hour lung preservation period, lungs were collected for proteomic profiling.