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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Presented here is a protocol to provide a step-by-step ischemia-free liver transplantation protocol under ex situ normothermic machine perfusion (37 °C) of human livers from donors to recipients.

Abstract

Currently, ex situ machine perfusion is a burgeoning technique that provides a better preservation method for donor organs than conventional static cold preservation (0–4 °C). A continuous blood supply to organs using machine perfusion from procurement and preservation to implantation facilitates complete prevention of ischemia reperfusion injury and permits ex situ functional assessment of donor livers before transplantation. In this manuscript, we provide a step-by-step ischemia-free liver transplantation protocol in which an ex situ normothermic machine perfusion apparatus is used for pulsatile perfusion through the hepatic artery and continuous perfusion of the portal vein from human donor livers to recipients. In the perfusion period, biochemical analysis of the perfusate is conducted to assess the metabolic activity of the liver, and a liver biopsy is also performed to evaluate the degree of injury. Ischemia-free liver transplantation is a promising method to avoid ischemia-reperfusion injury and may potentially increase the donor pool for transplantation.

Introduction

Ischemia reperfusion injury (IRI) is a well-known and widespread complication in organ transplantation. Obvious nonimmunological events lead to poor graft outcomes and delayed graft function, which are related to the high proportions of organ failure, re-transplantation, and recipient death1. Conventional cold storage (CCS) of organs was previously identified as a classic method to slow down metabolism but it does not have an influence on preventing progressive dysfunction and damage to cellular integrity. Furthermore, leukocyte accumulation is induced by reactive oxygen metabolites in the reperfusion phase. All of these biological processes be....

Protocol

This protocol was reviewed and approved by the ethics committee of The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China. Informed consent was obtained from all the participants. All the procedures in studies involving human participants were performed in accordance with the 1964 Helsinki Declaration and its later amendments or revisions.

1. Preparation of the perfusion solution and device

NOTE: The total volume of the perfusion solution prepared f.......

Representative Results

In April 2018, a 66-year-old male donor with brain death was not considered by local transplant centers because of the high risk of graft loss in such donors. The reasons for discarding the liver, at the time of procurement were older age and macroscopic appearance of moderate firmness, round liver edges and suboptimal liver graft perfusion along with major donor comorbidities, which included hypertension, hypertensive heart disease, and the following associated factors: hypernatremia (sodium, 156 mmol/L) and hemodynamic.......

Discussion

This IFLT technique was established to completely avoid IRI. This article provides a step-by-step IFLT protocol from organ procurement, ex situ preservation to implantation.

Based on NMP, IFLT provides an uninterrupted supply of blood and oxygen to grafts from procurement and perseveration to implantation. Numerous studies have shown that NMP has significant advantages in reducing IRI, improving organ viability, and repairing graft damage compared to static cold preservation1.......

Acknowledgements

This study was supported by the National Natural Science Foundation of China (81401324 and 81770410), Guangdong Provincial International Cooperation Base of Science and Technology (Organ Transplantation) (2015B050501002), Guangdong Provincial Natural Science Funds for Distinguished Young Scholars (2015A030306025), Special Support Program for Training High-Level Talent in Guangdong Province (2015TQ01R168), Pearl River Nova Program of Guangzhou (201506010014), and Scientific Program for Young Teachers of Sun Yat-sen University (16ykpy05), China.

....

Materials

NameCompanyCatalog NumberComments
10% calcium gluconateHebei Tiancheng Pharmaceutical Co, Ltd1S18112410130 mL
25% magnesium sulphateHebei Tiancheng Pharmaceutical Co, LtdH200338613 mL
5% sodium bicarbonateHuiyinbi Group Jiangxi Dongya Pharmaceutical Co, LtdH36020283The amount depends on the pH
Cefoperazone sodium and sulbactam sodiumPfizerH200205971.5 g
Compound Amino Acid InjectionGuangdong Litai Pharmaceutical Co., LtdH20063797250 mL
Crossed-matched leucocyte-depleted washed red cellsGuangzhou Blood CenterH200337391300 mL
HeparinChengdu Hepatunn Pharmaceutical Co., LtdH5102120937500 U
Liver AssistOrgan AssistOA.Li.Li.140Perfusion device
Liver Assist disposable packageOrgan AssistOA.Li.DP.540Disposable set and cannulas
MetronidazoleShanghai Baxter Healthcare Co., Ltd.H200033010.5 g
scalp acupunctureWuhan W.E.O.Science & Technology Development Co., LtdWEO-JX-32B-5.0 0.7*25mmBile duct cannula
Succinylated gelatinorB. Braun Medical Suzhou Co., LtdH201131191400 mL

References

  1. Hanidziar, D., Koulmanda, M. Towards cytoprotection in the peritransplant period. Seminars in Immunology. 23 (3), 209-213 (2011).
  2. Eltzschig, H. K., Eckle, T. Ischemia and reperfusion-from mechanism to translation.

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Ischemia Reperfusion InjuryLiver TransplantationEx Situ Machine PerfusionStatic Cold PreservationNormothermic Machine PerfusionHepatic ArteryPortal VeinPerfusate AnalysisLiver BiopsyDonor Pool

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