title: Organotypic lung culture: A new model for studying ischemia and ex vivo perfusion in lung transplantation.
abstract: OBJECTIVE
  Donors after cardiac death (DCD) in lung transplantation is considered as a solution for organ shortage. However, it is characterized by warm ischemic period, which could be involved in severe Ischemia-Reperfusion lesion (IR) with early graft dysfunction. We describe a new hybrid model combining in vivo ischemia followed by in vitro reoxygenation using organ-specific culture.
  
  
  METHODS
  A hybrid model using in vivo ischemic period followed by in vitro lung slice reoxygenation was set up in rat to mimic DCD in lung transplantation with in vitro perfusion. Different markers (bioenergetics, oxidant stress assays, and histology) were measured to evaluate the viability of lung tissue after different ischemic times (I-0, I-1, I-2, I-4, I-15 hours) and reoxygenation times (R-0, R-1, R-4, R-24 hours).
  
  
  RESULTS
  No differences were found in cell viability, ATP concentrations, extracellular LDH assays or histology, demonstrating extensive viability of up to 4 hours in lung tissue warm ischemia. We found oxidative stress mainly during the ischemic period with no burst at reoxygenation. Cytosolic anti-oxidant system was involved first (I-0,I-1,I-2) followed by mitochondrial anti-oxidant system for extensive ischemia (I-4). Histological features showed differences in this model of ischemia-reoxygenation between bronchial epithelium and lung parenchymal cells, with epithelium regeneration after 2 hours of warm ischemia and 24 hours of perfusion.
  
  
  CONCLUSIONS
  The results of our hybrid model experiment suggest extensive lung viability of up to 4 hours ischemia. Our model could be an interesting tool to evaluate ex vivo reconditioning techniques after different in vivo lung insults.
authors.affiliation: a Department of General and Thoracic Surgery , Rouen University Hospital , France.|b Inserm U1096, Rouen University , France.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Baste
authors.affiliation: c Department of Thoracic and Cardio-Vascular Surgery , Rouen University Hospital , France.|d ABTE Toxemac EA 4651, Rouen University , France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Gay
authors.affiliation: c Department of Thoracic and Cardio-Vascular Surgery , Rouen University Hospital , France.|d ABTE Toxemac EA 4651, Rouen University , France.
authors.forename: Hassiba
authors.initials: H
authors.lastname: Smail
authors.affiliation: d ABTE Toxemac EA 4651, Rouen University , France.
authors.forename: Romain
authors.initials: R
authors.lastname: Noël
authors.affiliation: e Unit of Biostatistics , Rouen University Hospital , France.
authors.forename: Michael
authors.initials: M
authors.lastname: Bubenheim
authors.affiliation: f Department of Pathology , Bordeaux University Hospital , France.
authors.forename: Hugues
authors.initials: H
authors.lastname: Begueret
authors.affiliation: d ABTE Toxemac EA 4651, Rouen University , France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Morin
authors.affiliation: b Inserm U1096, Rouen University , France.|c Department of Thoracic and Cardio-Vascular Surgery , Rouen University Hospital , France.
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Litzler
mesh_terms: D000818:Animals; D004734:Energy Metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D008168:Lung / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D016040:Lung Transplantation; D008297:Male; D009924:Organ Culture Techniques; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D052096:Warm Ischemia
chemical_list: D017382:Reactive Oxygen Species; D005979:Glutathione Peroxidase
keywords: ex vivo lung perfusion; ischemia-reoxygenation; lung transplantation; organotypic culture; warm ischemia