Liver transplantation remains to be the only chance for survival in a variety of different diseases leading to acute or chronic hepatic failure. Since its first successful application in mankind in 1963 by Thomas E. Starzl, the concept of liver transplantation has evolved into a reliable treatment option applied worldwide, mainly as a result of advancements in the understanding of the immune system, the development of modern immunosuppression, and the optimization of perioperative care and surgical techniques1,2. However, aging populations and a higher demand for organs have resulted in donor shortages, with increased use of marginal grafts from extended criteria donors and the emergence of new challenges in the past decades. The introduction and widespread implementation of organ machine perfusion is believed to open up an array of possibilities with regard to graft reconditioning and modulation and to help mitigate organ shortages and reduce waiting list mortality3,4,5,6.
In order to evaluate these concepts and their effects in vivo, translational transplant models are necessary7. In 1983, Kamada et al. introduced an efficient orthotopic liver transplant model in rats that has since been extensively modified and applied by workgroups around the globe8,9,10,11. The orthotopic liver transplant model in mice is technically more demanding, but also more valuable in terms of immunological transferability, and was first reported in 1991 by Qian et al.12. Despite advantages regarding availability, animal welfare, and costs, rodent models are limited in their applicability in clinical settings7. Hence, large animal models are required.
In recent years, pigs have become the main animal species used for translational research due to their anatomical and physiological similarities with humans. Furthermore, current progress in the field of xenotransplantation might further increase the importance of pigs as research objects13,14.
Garnier et al. described a liver transplant model in pigs as early as 196515. Several authors, including Calne et al. in 1967 and Chalstrey et al. in 1971, subsequently reported modifications, ultimately leading to a safe and feasible concept of experimental porcine liver transplantation in the decades to follow16,17,18,19,20,21.
More recently, different work groups have provided data with regard to current issues in liver transplantation using a technique of porcine orthotopic liver transplantation, almost invariably including an active or passive veno-venous, i.e., porto-caval, bypass19,22. The reason for this is a species-specific intolerance to the clamping of the vena cava inferior and the portal vein during the anhepatic phase due to a comparatively larger intestine and fewer porto-caval or cavo-caval shunts (e.g., lack of a vena azygos), resulting in increased perioperative morbidity and mortality23. Vena cava inferior-sparing transplant techniques applied in human recipients as an alternative are not feasible as the porcine vena cava inferior is encased by hepatic tissue23.
However, the usage of a veno-venous bypass further increases technical and logistical complexity in an already demanding surgical procedure, therefore possibly preventing workgroups from attempting implementation of the model altogether. Apart from the direct physiological and immunological effects of a bypass, some authors have pointed out the significant morbidity such as blood loss or air embolism during shunt placement and the need for a simultaneous splenectomy, potentially affecting short- and long-term results after engraftment24,25.
The following protocol describes a simple technique of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h, representing extended criteria donor conditions without the usage of a veno-venous bypass during engraftment, including donor liver procurement, back-table preparation, recipient hepatectomy, and anesthesiological pre- and intraoperative management.
This model should be of special interest for surgical workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, the reconditioning of extended criteria donor organs, and associated immunological mechanisms.