Learning curves.

<p>(A) Surgical preparation time (from heparin administration to flushing; R = 0.291, p = 0.397). (B) Procurement time (from flushing to harvest; R = 0.660, p = 0.0003); (C) Connection time (from harvest to lung connection to the circuit; R = 0.299, p = 0.370). Nonlinear regression analysis.</p

Ex Vivo Lung Perfusion in the Rat: Detailed Procedure and Videos

<div><p>Ex vivo lung perfusion (EVLP) is a promising procedure for evaluation, reconditioning, and treatment of marginal lungs before transplantation. Small animal models can contribute to improve clinical development of this technique and represent a substantial platform for bio-molecular investigations. However, to accomplish this purpose, EVLP models must sustain a prolonged reperfusion without pharmacological interventions. Currently available protocols only partly satisfy this need. The aim of the present research was accomplishment and optimization of a reproducible model for a protracted rat EVLP in the absence of anti-inflammatory treatment. A 180 min, uninjured and untreated perfusion was achieved through a stepwise implementation of the protocol. Flow rate, temperature, and tidal volume were gradually increased during the initial reperfusion phase to reduce hemodynamic and oxidative stress. Low flow rate combined with open atrium and protective ventilation strategy were applied to prevent lung damage. The videos enclosed show management of the most critical technical steps. The stability and reproducibility of the present procedure were confirmed by lung function evaluation and edema assessment. The meticulous description of the protocol provided in this paper can enable other laboratories to reproduce it effortlessly, supporting research in the EVLP field.</p></div

Reagents employed.

<p>Reagents employed.</p

General protocol features and perfusion fluid characteristics of recently reported EVLP rat models.

<p>General protocol features and perfusion fluid characteristics of recently reported EVLP rat models.</p

Instruments employed.

<p>Instruments employed.</p

Comparison of reconditioning phase settings in recently reported rat EVLP protocols.

<p>Comparison of reconditioning phase settings in recently reported rat EVLP protocols.</p

Changes over time in perfusion fluid parameters.

<p>Changes over time in perfusion fluid parameters.</p

Comparison of reperfusion phase settings in recently reported rat EVLP protocols.

<p>Comparison of reperfusion phase settings in recently reported rat EVLP protocols.</p

Lung parameter changes during EVLP.

<p>(A) Pulmonary artery pressure (PAP). (B) Total pulmonary vascular resistance (TPVR). (C) Peak inspiratory pressure (Ppeak). (D) Dynamic compliance (Cdyn). Pressure recruitment maneuvers (RMs) were performed at 40 and 175 min. Results are expressed as the mean ± SEM (N = 10). One-way repeated measures ANOVA; p value was <0.001 for each parameter considered. Tukey’s multiple comparison test.</p

Protocol timeline.

<p>Schematic overview of EVLP protocol. Abbreviations: VT, tidal volume; RR, respiratory rate; PEEP, positive end expiratory pressure.</p