Original Pre-Clinical ScienceSustained function of genetically modified porcine lungs in an ex vivo model of pulmonary xenotransplantation
Section snippets
Generation of transgenic pigs
GTKO pigs were generated by homologous recombination and cloning by nuclear transfer, as previously described.5 These pigs lack expression of the Gal epitope on all organs and tissues but are otherwise phenotypically normal. Transgenic pigs coexpressing the human complement regulators CD55 and CD59 were generated by conventional oocyte microinjection.6 Transgenic pigs expressing human CD39 were produced by transfection of pig fetal fibroblasts followed by cloning using stable transfectants as
Establishing the ex vivo rig
For the 10 experiments, the mean time from instillation of pneumoplegia to lung explant and ice preservation was 20 ± 2 minutes. Cannulation to the ex vivo rig and initial perfusion with Steen solution took a further 88 ± 2 minutes, after which it took 74 ± 4 minutes to establish stable physiologic ventilator and perfusion function. Wild-type porcine lungs demonstrated stable physiologic function for 6 hours while being continuously perfused with Steen solution (data not shown).
Lung survival and performance
Wild-type lungs
Discussion
In this series of preliminary experiments on ex vivo performance of porcine lungs after addition of human blood, we have demonstrated that genetic modification of the donor results in superior graft function compared with wild-type lungs. Specifically, we were able to demonstrate superior oxygenation of the perfusate and reduced pulmonary vascular resistance when human blood was perfused through GTKO, GTKO/CD55-59 and GTKO/CD55-59/CD39 lungs compared with wild-type porcine lungs. Although there
Disclosure statement
The authors have no conflicts of interest to disclose.
This study was supported by a Norman E. Shumway International Society for Heart and Lung Transplantation (ISHLT) Career Development Award (to G.W.) and by the Margaret Pratt Foundation.
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