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Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls

Porcine models of liver normothermic machine perfusion (NMP) are increasingly used in transplant research, although known to be challenging because of their complex methodology and their scarcely documented operational aspects. Here, we aimed to provide a methodological framework for researchers loo...

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Autores principales: Gilbo, Nicholas, Wylin, Tine, Heedfeld, Veerle, Jochmans, Ina, Pirenne, Jacques, Friend, Peter, Monbaliu, Diethard
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Lippincott Williams & Wilkins 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8670579/
https://www.ncbi.nlm.nih.gov/pubmed/34912950
http://dx.doi.org/10.1097/TXD.0000000000001276
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author Gilbo, Nicholas
Wylin, Tine
Heedfeld, Veerle
Jochmans, Ina
Pirenne, Jacques
Friend, Peter
Monbaliu, Diethard
author_facet Gilbo, Nicholas
Wylin, Tine
Heedfeld, Veerle
Jochmans, Ina
Pirenne, Jacques
Friend, Peter
Monbaliu, Diethard
author_sort Gilbo, Nicholas
collection PubMed
description Porcine models of liver normothermic machine perfusion (NMP) are increasingly used in transplant research, although known to be challenging because of their complex methodology and their scarcely documented operational aspects. Here, we aimed to provide a methodological framework for researchers looking to adopt NMP technology in research setting by giving an in-detail account of the implementation of a previously validated porcine liver NMP model. We subjected groups of 3–5 porcine livers to 24 h NMP and, using a trial-and-error principle, introduced stepwise changes in the NMP setting with the objective to obtain stable preservation of liver function and histology for 24 h. Female porcine livers were procured, and packed red-blood-cell perfusate was prepared. Perfusate oxygenation, hemodynamics, markers of hepatic injury (aspartate transaminase [AST]), function (lactate, perfusate pH, bile production), and histology were analyzed. Intermediate analysis was performed within groups and a minimum of 3 (out of 5) failed experiments prompted methodological reevaluation. Overall, 13 liver NMP experiments were needed in 3 phases. In phase 1, loss of oxygenator performance occurred from 6 h onward in 3 consecutive experiments because of perfusate leakage. In phase 2, a plasma-tight hollow fiber oxygenator ensured adequate perfusate oxygenation in 5 experiments. However, portal vein resistance increased during all liver NMP, associated with high perfusate AST levels (range, 106–322 IU/L/100 g) and pan-lobular sinusoidal dilation and hemorrhage, suggesting liver outflow impairment. In phase 3, an improved inferior vena cava cannulation technique avoided liver outflow impairment, resulting in lower AST release (range, 29–101 IU/L/100 g), improved lactate clearance, preserved biliary excretion, and normal histology in 5 experiments. This study underscores the critical importance of auditing all equipment and operational components of NMP circuits to obtain successful and reproducible perfusion setup and advocates for in-detail reporting of methodological aspects and potential pitfalls.
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spelling pubmed-86705792021-12-14 Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls Gilbo, Nicholas Wylin, Tine Heedfeld, Veerle Jochmans, Ina Pirenne, Jacques Friend, Peter Monbaliu, Diethard Transplant Direct Liver Transplantation Porcine models of liver normothermic machine perfusion (NMP) are increasingly used in transplant research, although known to be challenging because of their complex methodology and their scarcely documented operational aspects. Here, we aimed to provide a methodological framework for researchers looking to adopt NMP technology in research setting by giving an in-detail account of the implementation of a previously validated porcine liver NMP model. We subjected groups of 3–5 porcine livers to 24 h NMP and, using a trial-and-error principle, introduced stepwise changes in the NMP setting with the objective to obtain stable preservation of liver function and histology for 24 h. Female porcine livers were procured, and packed red-blood-cell perfusate was prepared. Perfusate oxygenation, hemodynamics, markers of hepatic injury (aspartate transaminase [AST]), function (lactate, perfusate pH, bile production), and histology were analyzed. Intermediate analysis was performed within groups and a minimum of 3 (out of 5) failed experiments prompted methodological reevaluation. Overall, 13 liver NMP experiments were needed in 3 phases. In phase 1, loss of oxygenator performance occurred from 6 h onward in 3 consecutive experiments because of perfusate leakage. In phase 2, a plasma-tight hollow fiber oxygenator ensured adequate perfusate oxygenation in 5 experiments. However, portal vein resistance increased during all liver NMP, associated with high perfusate AST levels (range, 106–322 IU/L/100 g) and pan-lobular sinusoidal dilation and hemorrhage, suggesting liver outflow impairment. In phase 3, an improved inferior vena cava cannulation technique avoided liver outflow impairment, resulting in lower AST release (range, 29–101 IU/L/100 g), improved lactate clearance, preserved biliary excretion, and normal histology in 5 experiments. This study underscores the critical importance of auditing all equipment and operational components of NMP circuits to obtain successful and reproducible perfusion setup and advocates for in-detail reporting of methodological aspects and potential pitfalls. Lippincott Williams & Wilkins 2021-12-13 /pmc/articles/PMC8670579/ /pubmed/34912950 http://dx.doi.org/10.1097/TXD.0000000000001276 Text en Copyright © 2021 The Author(s). Transplantation Direct. Published by Wolters Kluwer Health, Inc. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) , where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.
spellingShingle Liver Transplantation
Gilbo, Nicholas
Wylin, Tine
Heedfeld, Veerle
Jochmans, Ina
Pirenne, Jacques
Friend, Peter
Monbaliu, Diethard
Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title_full Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title_fullStr Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title_full_unstemmed Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title_short Porcine Liver Normothermic Machine Perfusion: Methodological Framework and Potential Pitfalls
title_sort porcine liver normothermic machine perfusion: methodological framework and potential pitfalls
topic Liver Transplantation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8670579/
https://www.ncbi.nlm.nih.gov/pubmed/34912950
http://dx.doi.org/10.1097/TXD.0000000000001276
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