Background
The clinical application of normothermic ex vivo lung perfusion (EVLP) has increased
donor lung utilization for transplantation through functional assessment. To develop
it as a platform for donor lung repair, reconditioning and regeneration, the perfusate
should be modified to support the lung during extended EVLP.
Methods
Human lung epithelial cells and pulmonary microvascular endothelial cells were cultured,
and the effects of Steen solution (commonly used EVLP perfusate) on basic cellular
function were tested. Steen solution was modified based on screening tests in cell
culture, and further tested with an EVLP cell culture model, on apoptosis, GSH, HSP70,
and IL-8 expression. Finally, a modified formula was tested on porcine EVLP. Physiological
parameters of lung function, histology of lung tissue, and amino acid concentrations
in EVLP perfusate were measured.
Results
Steen solution reduced cell confluence, induced apoptosis, and inhibited cell migration,
compared to regular cell culture media. Adding L-alanyl-L-glutamine to Steen solution
improved cell migration and decreased apoptosis. It also reduced cold preservation
and warm perfusion-induced apoptosis, enhanced GSH and HSP70 production, and inhibited
IL-8 expression on an EVLP cell culture model. L-alanyl-L-glutamine modified Steen
solution supported porcine lungs on EVLP with significantly improved lung function,
well-preserved histological structure, and significantly higher levels of multiple
amino acids in EVLP perfusate.
Conclusions
Adding L-alanyl-L-glutamine to perfusate may provide additional energy support, antioxidant,
and cytoprotective effects to lung tissue. The pipeline developed herein, with cell
culture, cell EVLP, and porcine EVLP models, can be used to further optimize perfusates
to improve EVLP outcomes.
Keywords
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Article info
Publication history
Published online: November 03, 2022
Footnotes
Tweeter Handle: Ex vivo lung perfusion (EVLP) has been used in donor lung assessment and developed as a platform for donor lung repair. A pipeline was developed with cell culture, cell EVLP, and porcine EVLP models. Adding L-alanyl-L-glutamine to EVLP perfusate provides additional energy support, antioxidant, and cytoprotective effects to lung tissue. It may improve EVLP outcomes for donor lung repair, reconditioning and bioengineering.
Identification
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© 2022 International Society for Heart and Lung Transplantation. All rights reserved.
