Technique for ex vivo lung perfusion (EVLP) of lungs from brain-dead donor rats and testing the effect of prednisolone treatment

J.E. van Zanden, H.G.D. Leuvenink, E.A.M. Verschuuren, S. Veldhuis, P. Ottens, M.E. Erasmus, C.M.V. Hottenrott

Chair(s): dr. Martin. J. Hoogduijn, wetenschappelijk medewerker, Erasmus MC & prof. dr. Cees. van Kooten, onderzoeker, LUMC

Wednesday 9 march 2016

16:20 - 16:30h at Zaal 5 & 6

Categories: Parallelsessie (basaal)

Parallel session: Parallelsessie IV - Basaal

Ex vivo lung perfusion (EVLP) systems have become an important tool to treat marginal brain dead donor lungs with pulmonary edema. Some centers have successfully increased their donor lung pool using EVLP while graft survival has remained comparable to standard brain dead donor lung transplantation. This is probably the result of a comparable immune response after transplantation. Therefore, an easily reproducible and stable rat lung perfusion system is an important tool to test new treatment opportunities to limit the immune response and possibly support the regeneration of donor lungs. Heart-lung blocks were procured from Lewis rats three hours after acute traumatic brain death induction, and were subsequently cold preserved for one hour. Thereafter, lungs were placed for six hours in the normothermic EVLP model. Lungs were ventilated with a pressure controlled ventilation adjusted to a tidal volume (VT) of 7 ml/kg of body weight, a PEEP of 5 cmH2O, a frequency of 60 and a FiO2 of 21%. Perfusion was performed with a modified Steen solution and cefuroxime at a maximal pulmonary arterial pressure of 13 mmHg. As intervention we study the effect of prednisolone (40 mg), standardly administered in the human EVLP. Ventilation parameters, lung oxygenation capacity, pH, glucose levels and flow were noted and perfusate samples collected, over time. Lungs were macroscopically scored and tissue was stored for wet/dry ratio, qPCR and patho-histological analysis. Judged by ventilation parameters, lung oxygenation capacity and flow, a stable rat EVLP system was developed. Preliminary analysis suggest that addition of prednisolone in the EVLP system results in a better macroscopic appearance, ventilation parameters and lung oxygenation capacity. We hope to also soon be able to present the effect of prednisolone on immune activation in the lung. This model will be an important tool to test new treatment opportunities for brain death induced immune activation of donor lungs. Since prednisolone is known to have an anti-inflammatory effect, it might be beneficial to exclude it when testing future treatment opportunities in the rat EVLP.