Step-by-Step Guide for a Simplified Model of Porcine Orthotopic Liver Transplant

Ischemic-type biliary lesions (ITBLs) arise most frequently after donation after circulatory death (DCD) liver transplantation and result in high morbidity and graft loss. Many DCD grafts are discarded out of fear for this complication. In theory, microvascular thrombi deposited during donor warm ischemia might be implicated in ITBL pathogenesis. Herein, we aim to evaluate the effects of the administration of either heparin or the fibrinolytic drug tissue plasminogen activator (TPA) as means to improve DCD liver graft quality and potentially avoid ITBL. Donor pigs were subjected to 1 hour of cardiac arrest (CA) and divided among 3 groups: no pre-arrest heparinization nor TPA during postmortem regional perfusion; no pre-arrest heparinization but TPA given during regional perfusion; and pre-arrest heparinization but no TPA during regional perfusion. In liver tissue sampled 1 hour after CA, fibrin deposition was not detected, even when heparin was not given prior to arrest. Although it was not useful to prevent microvascular clot formation, pre-arrest heparin did offer cytoprotective effects during CA and beyond, reflected in improved flows during regional perfusion and better biochemical, functional, and histological parameters during posttransplantation follow-up. In conclusion, this study demonstrates the lack of impact of TPA use in porcine DCD liver transplantation and adds to the controversy over whether the use of TPA in human DCD liver transplantation really offers any protective effect. On the other hand, when it is administered prior to CA, heparin does offer anti-inflammatory and other cytoprotective effects that help improve DCD liver graft quality. Liver Transplantation 24 665-676 2018 AASLD.

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“…Donor anesthesia was performed as described previously . The liver and hilar structures were dissected, and cholecystectomy was performed.…”

Section: Methodsmentioning

confidence: 99%

“…Donor anesthesia was performed as described previously. (14) The liver and hilar structures were dissected, and cholecystectomy was performed. Hepatic artery flow (HAF) and portal vein flow (PVF) were measured using ultrasonic flow probes connected to a flowmeter (HT107, Transonic Systems, Ithaca, NY).…”

Section: Donor Procedures and Nrpmentioning

confidence: 99%

Heparin but not tissue plasminogen activator improves outcomes in donation after circulatory death liver transplantation in a porcine model

et al. 2018

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“…Surgical procedures, intraoperative monitoring and postoperative management SFS liver transplants were performed as described previously (4,18). Briefly, donor pigs (15-20 kg) underwent extended hepatectomy.…”

Section: Animalsmentioning

confidence: 99%

Somatostatin Therapy Protects Porcine Livers in Small-for-Size Liver Transplantation

Hessheimer

Escobar

Muñoz

et al. 2014

American Journal of Transplantation

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Small-for-size (SFS) injury occurs in partial liver transplantation due to several factors, including excessive portal inflow and insufficient intragraft responses. We aim to determine the role somatostatin plays in reducing portal hyperperfusion and preventing the cascade of deleterious events produced in small grafts. A porcine model of 20% liver transplantation is performed. Perioperatively treated recipients receive somatostatin and untreated controls standard intravenous fluids. Recipients are followed for up to 5 days. In vitro studies are also performed to determine direct protective effects of somatostatin on hepatic stellate cells (HSC) and sinusoidal endothelial cells (SEC). At reperfusion, portal vein flow (PVF) per gram of tissue increased fourfold in untreated animals versus approximately threefold among treated recipients (p ¼ 0.033). Postoperatively, markers of hepatocellular, SEC and HSC injury were improved among treated animals. Hepatic regeneration occurred in a slower but more orderly fashion among treated grafts; functional recovery was also significantly better. In vitro studies revealed that somatostatin directly reduces HSC activation, though no direct effect on SEC was found. In SFS transplantation, somatostatin reduces PVF and protects SEC in the critical postreperfusion period. Somatostatin also exerts a direct cytoprotective effect on HSC, independent of changes in PVF.

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“…Among all the available animal models, pigs serve as the most clinically relevant model for human liver disease, given the similarities between pig liver and human liver in morphology, structure and physiology [7], [8], [9]. In 2009, iPS cells were reprogrammed from pig primary ear fibroblasts using four reprogramming genes including Sox2, Klf4, Oct4, and c-Myc [10].…”

Section: Introductionmentioning

confidence: 99%

High Efficient Differentiation of Functional Hepatocytes from Porcine Induced Pluripotent Stem Cells

Mich-Basso

Lin

et al. 2014

PLoS ONE

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Hepatocyte transplantation is considered to be a promising therapy for patients with liver diseases. Induced pluripotent stem cells (iPSCs) provide an unlimited source for the generation of functional hepatocytes. In this study, we generated iPSCs from porcine ear fibroblasts (PEFs) by overexpressing Sox2, Klf4, Oct4, and c-Myc (SKOM), and developed a novel strategy for the efficient differentiation of hepatocyte-like cells from porcine iPSCs by following the processes of early liver development. The differentiated cells displayed the phenotypes of hepatocytes, exhibited classic hepatocyte-associated bio-functions, such as LDL uptake, glycogen storage and urea secretion, as well as possessed the metabolic activities of cytochrome P-450 (CYP) 3A and 2C. Furthermore, we compared the hepatocyte differentiation efficacy of our protocol with another published method, and the results demonstrated that our differentiation strategy could significantly improve the generation of morphological and functional hepatocyte-like cells from porcine iPSCs. In conclusion, this study establishes an efficient method for in vitro generation of functional hepatocytes from porcine iPSCs, which could represent a promising cell source for preclinical testing of cell-based therapeutics for liver failure and for pharmacological applications.

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Step-by-Step Guide for a Simplified Model of Porcine Orthotopic Liver Transplant

Cited by 22 publications

References 18 publications

Heparin but not tissue plasminogen activator improves outcomes in donation after circulatory death liver transplantation in a porcine model

Heparin but not tissue plasminogen activator improves outcomes in donation after circulatory death liver transplantation in a porcine model

Somatostatin Therapy Protects Porcine Livers in Small-for-Size Liver Transplantation

High Efficient Differentiation of Functional Hepatocytes from Porcine Induced Pluripotent Stem Cells

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