Name: porcine liver transplantation without veno-venous bypass as an extended criteria donor model
Uploaded: 2022-08-17T14:00:00
Description: Watch this Scientific Journal Video about Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model at JoVE.com

The authors thank Britta Trautewig, Corinna L&#246;bbert, Astrid Dinkel, and Ingrid Meder for their diligence and commitment. Furthermore, the authors thank Tom Figiel for producing the picture material.

This study was performed at the Laboratory for Animal Science of Hannover Medical School after approval by the Lower Saxony regional authority for consumer protection and food safety (Nieders&#228;chsisches Landesamt f&#252;r Verbraucherschutz und Lebensmittelsicherheit [LAVES]; 19/3146)
1. Donor liver procurement
NOTE: The liver donors were female domestic pigs (Sus scrofa domesticus), aged 4-5 months old and with an average body weight of a...

<ol>
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T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation:+Fifty+years+of+experience.">Liver transplantation: Fifty years of experience.</a> World Journal of Gastroenterology. 20 (18), 5363-5374 (2014).</li><li>Jakubauskas, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Machine+perfusion+in+liver+transplantation:+A+systematic+review+and+meta-analysis.">Machine perfusion in liver transplantation: A systematic review and meta-analysis.</a> Visceral Medicine. , (2021).</li><li>Serifis, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Machine+perfusion+of+the+liver:+A+review+of+clinical+trials.">Machine perfusion of the liver: A review of clinical trials.</a> Frontiers in Surgery. 8, 625394 (2021).</li><li>Ceresa, C. 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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Machine+perfusion+of+the+liver:+Applications+in+transplantation+and+beyond.">Machine perfusion of the liver: Applications in transplantation and beyond.</a> Nature Reviews Gastroenterology &amp; Hepatology. 19 (3), 199-209 (2022).</li><li>Schlegel, A., Muller, X., Dutkowski, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Machine+perfusion+strategies+in+liver+transplantation.">Machine perfusion strategies in liver transplantation.</a> Hepatobiliary Surgery and Nutrition. 8 (5), 490-501 (2019).</li><li>Wenzel, N., Blasczyk, R., Figueiredo, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+in+allogenic+solid+organ+transplantation.">Animal models in allogenic solid organ transplantation.</a> Transplantology. 2 (4), 412-424 (2021).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+surgical+experience+with+five+hundred+thirty+liver+transplants+in+the+rat.">A surgical experience with five hundred thirty liver transplants in the rat.</a> Surgery. 93 (1), 64-69 (1983).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> Journal of Visualized Experiments. (65), e4143 (2012).</li><li>Yang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+model+of+orthotopic+liver+transplantation+using+a+novel+magnetic+anastomosis+technique+for+suprahepatic+vena+cava+reconstruction.">A rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction.</a> Journal of Visualized Experiments. (133), e56933 (2018).</li><li>Chen, X. -. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+complications+after+arterial+reconnection+in+a+rat+model+of+orthotopic+liver+transplantation.">Reduced complications after arterial reconnection in a rat model of orthotopic liver transplantation.</a> Journal of Visualized Experiments. (165), e60628 (2020).</li><li>Qian, S. G., Fung, J. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+mouse.">Orthotopic liver transplantation in the mouse.</a> Transplantation. 52 (3), 562-564 (1991).</li><li>Li, X., Wang, Y., Yang, H., Dai, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+and+hepatocyte+transplantation:+What+can+pigs+contribute.">Liver and hepatocyte transplantation: What can pigs contribute.</a> Frontiers in Immunology. 12, 802692 (2022).</li><li>Reardon, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+pig-to-human+heart+transplant:+what+can+scientists+learn.">First pig-to-human heart transplant: what can scientists learn.</a> Nature. 601 (7893), 305-306 (2022).</li><li>Garnier, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+the+pig:+Surgical+approach.">Liver transplantation in the pig: Surgical approach.</a> Comptes Rendus Hebdomadaires des Seances de l'Academie des Sciences. Serie d: Sciences Naturelles. 260 (21), 5621-5623 (1965).</li><li>Calne, R. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observations+of+orthotopic+liver+transplantation+in+the+pig.">Observations of orthotopic liver transplantation in the pig.</a> British Medical Journal. 2 (5550), 478-480 (1967).</li><li>Chalstrey, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+of+orthotopic+liver+transplantation+in+the+pig.">Technique of orthotopic liver transplantation in the pig.</a> The British Journal of Surgery. 58 (8), 585-588 (1971).</li><li>Filipponi, F., Falcini, F., Benassai, C., Martini, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplant+in+pigs:+Several+variations+of+the+surgical+technic.">Orthotopic liver transplant in pigs: Several variations of the surgical technic.</a> Il Giornale di Chirurgia. 10 (7-8), 374-378 (1989).</li><li>Spetzler, V. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+of+porcine+liver+procurement+and+orthotopic+transplantation+using+an+active+porto-caval+shunt.">Technique of porcine liver procurement and orthotopic transplantation using an active porto-caval shunt.</a> Journal of Visualized Experiments. (99), e52055 (2015).</li><li>Oldhafer, K. J., Hauss, J., Gubernatis, G., Pichlmayr, R., Spiegel, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+pigs:+A+model+for+studying+reperfusion+injury.">Liver transplantation in pigs: A model for studying reperfusion injury.</a> Journal of Investigative Surgery. 6 (5), 439-450 (1993).</li><li>Oldhafer, K. 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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+experimental+model+of+large-for-size+liver+transplantation+in+pigs.">A simplified experimental model of large-for-size liver transplantation in pigs.</a> Clinics. 68 (8), 1152-1156 (2013).</li><li>Schiefer, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+histamine+and+diamine+oxidase+in+patients+undergoing+orthotopic+liver+transplantation.">Regulation of histamine and diamine oxidase in patients undergoing orthotopic liver transplantation.</a> Scientific Reports. 10 (1), 822 (2020).</li><li>Esmaeilzadeh, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+guidelines+for+porcine+liver+allo-transplantation:+A+review+of+literature.">Technical guidelines for porcine liver allo-transplantation: A review of literature.</a> Annals of Transplantation. 17 (2), 101-110 (2012).</li><li>Oldhafer, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supportive+hepatocyte+transplantation+after+partial+hepatectomy+enhances+liver+regeneration+in+a+preclinical+pig+model.">Supportive hepatocyte transplantation after partial hepatectomy enhances liver regeneration in a preclinical pig model.</a> European Surgical Research. 62 (4), 238-247 (2021).</li><li>Stockmann, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+LiMAx+test:+A+new+liver+function+test+for+predicting+postoperative+outcome+in+liver+surgery.">The LiMAx test: A new liver function test for predicting postoperative outcome in liver surgery.</a> HPB. 12 (2), 139-146 (2010).</li><li>Lapisatepun, W., Lapisatepun, W., Agopian, V., Xia, V. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Venovenous+bypass+during+liver+transplantation:+A+new+look+at+an+old+technique.">Venovenous bypass during liver transplantation: A new look at an old technique.</a> Transplantation Proceedings. 52 (3), 905-909 (2020).</li><li>Falcini, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Veno-venous+bypass+in+experimental+liver+transplantation:+portal-jugular+versus+caval-portal-jugular.">Veno-venous bypass in experimental liver transplantation: portal-jugular versus caval-portal-jugular.</a> Il Giornale di Chirurgia. 11 (4), 206-210 (1990).</li><li>Copca, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+liver+transplantation+on+pigs+--+Technical+considerations.">Experimental liver transplantation on pigs -- Technical considerations.</a> Chirurgia. 108 (4), 542-546 (2013).</li><li>Torres, O. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamic+alterations+during+orthotopic+liver+experimental+transplantation+in+pigs.">Hemodynamic alterations during orthotopic liver experimental transplantation in pigs.</a> Acta Cirurgica Brasileria. 23 (2), 135-139 (2008).</li><li>Canedo, B. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+autotransplantation+in+pigs+without+venovenous+bypass:+A+simplified+model+using+a+supraceliac+aorta+cross-clamping+maneuver.">Liver autotransplantation in pigs without venovenous bypass: A simplified model using a supraceliac aorta cross-clamping maneuver.</a> Annals of Transplantation. 20, 320-326 (2015).</li><li>Battersby, C., Hickman, R., Saunders, S. J., Terblanche, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+function+in+the+pig.+1.+The+effects+of+30+minutes'+normothermic+ischaemia.">Liver function in the pig. 1. The effects of 30 minutes' normothermic ischaemia.</a> The British Journal of Surgery. 61 (1), 27-32 (1974).</li><li>Kaiser, G. M., Heuer, M. M., Fr&#252;hauf, N. R., K&#252;hne, C. A., Broelsch, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=General+handling+and+anesthesia+for+experimental+surgery+in+pigs.">General handling and anesthesia for experimental surgery in pigs.</a> Journal of Surgical Research. 130 (1), 73-79 (2006).</li><li>Oike, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+technique+of+orthotopic+liver+transplantation+in+pigs.">Simplified technique of orthotopic liver transplantation in pigs.</a> Transplantation. 71 (2), 328-331 (2001).</li><li>Heuer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+swine+without+venovenous+bypass.">Liver transplantation in swine without venovenous bypass.</a> European Surgical Research. 45 (1), 20-25 (2010).</li><li>Fondevila, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Step-by-step+guide+for+a+simplified+model+of+porcine+orthotopic+liver+transplant.">Step-by-step guide for a simplified model of porcine orthotopic liver transplant.</a> The Journal of Surgical Research. 167 (1), 39-45 (2011).</li></ol>

Recent technical developments such as the introduction of machine perfusion have the potential to revolutionize the field of liver transplantation. In order to translate graft reconditioning or modification concepts into clinical settings, reproducible transplant models in large animals are inevitable.
After the initial introduction of porcine orthotopic liver transplantation, several authors have worked on the improvement of these techniques over the past five&#160;decades. Differences within...

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.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Abdominal retractor</td>
			<td>No Company Name available</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Aortic clamp, straight</td>
			<td>Firma Martin</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Arterial Blood Sampler Aspirator (safePICOAspirator) 1.5 mL</td>
			<td>Radiometer Medical ApS</td>
			<td>956-622</td>
			<td></td>
		</tr>
		<tr>
			<td>Atropine (Atropinsulfat 0.5 mg/1 mL)</td>
			<td>B.Braun</td>
			<td>648037</td>
			<td></td>
		</tr>
		<tr>
			<td>Backhaus clamp</td>
			<td>Bernshausen</td>
			<td>BF432</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar forceps, 23 cm&#160;</td>
			<td>SUTTER</td>
			<td>780222 SG</td>
			<td></td>
		</tr>
		<tr>
			<td>Bowl 5 L, 6 L, 9 L</td>
			<td>Chiru-Instrumente</td>
			<td>35-114327</td>
			<td></td>
		</tr>
		<tr>
			<td>Braunol Braunoderm</td>
			<td>B.Braun</td>
			<td>3881059</td>
			<td></td>
		</tr>
		<tr>
			<td>Bulldog clamp</td>
			<td>Aesculap</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Button canula</td>
			<td>Krauth + Timmermann GmbH</td>
			<td>1464LL1B</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium gluconate (2.25 mmol/10 mL (10%))</td>
			<td>B.Braun</td>
			<td>2353745</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Saver (Autotransfusion Reservoir)</td>
			<td>Fresenius Kabi AG</td>
			<td>9108471</td>
			<td></td>
		</tr>
		<tr>
			<td>Central venous catheter 7Fr., 3 Lumina, 30 cm 0.81 mm</td>
			<td>Arrow</td>
			<td>AD-24703</td>
			<td></td>
		</tr>
		<tr>
			<td>Clamp</td>
			<td>INOX</td>
			<td>B-17845&#160; /&#160; BH110&#160; / B-481</td>
			<td></td>
		</tr>
		<tr>
			<td>Clamp</td>
			<td>Aesculap</td>
			<td>AN909R</td>
			<td></td>
		</tr>
		<tr>
			<td>Clamp, 260 mm</td>
			<td>Fehling Instruments GMbH &#38;Co.KG</td>
			<td>ZAU-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Clip Forceps, medium</td>
			<td>Ethicon</td>
			<td>LC207</td>
			<td></td>
		</tr>
		<tr>
			<td>Clip forceps, small</td>
			<td>Ethicon&#160;</td>
			<td>LC107</td>
			<td></td>
		</tr>
		<tr>
			<td>CPDA-1 solution</td>
			<td>Fresenius Kabi AG</td>
			<td>41SD09AA00</td>
			<td></td>
		</tr>
		<tr>
			<td>Custodiol (Histidin-Tryptophan-Ketogluterat-Solution)</td>
			<td>Dr.Franz K&#246;hler Chemie GmbH</td>
			<td>2125921</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting scissors</td>
			<td>LAWTON&#160; 05-0641&#160;</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting scissors, 180 mm</td>
			<td>Metzenbaum&#160;</td>
			<td>BC606R</td>
			<td></td>
		</tr>
		<tr>
			<td>Endotracheal tube 8.0 mm</td>
			<td>Covetrus</td>
			<td>800764</td>
			<td></td>
		</tr>
		<tr>
			<td>Epinephrine (Adrenalin 1:1000)</td>
			<td>InfectoPharm</td>
			<td>9508734</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcon Tubes 50ml</td>
			<td>Greiner&#160;</td>
			<td>227 261 L</td>
			<td></td>
		</tr>
		<tr>
			<td>Femoralis clamp</td>
			<td>Ulrich&#160;</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Fentanyl 0.1mg</td>
			<td>PanPharma</td>
			<td>00483</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, anatomical</td>
			<td>Martin</td>
			<td>12-100-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, anatomical, 250 mm</td>
			<td>Aesculap</td>
			<td>BD052R</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, anatomical, 250 mm</td>
			<td>Aesculap</td>
			<td>BD032R</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, anatomical, 250 mm&#160;</td>
			<td>Aesculap</td>
			<td>BD240R</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, surgical</td>
			<td>Bernshausen</td>
			<td>BD 671</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps, surgical</td>
			<td>INOX</td>
			<td>B-1357</td>
			<td></td>
		</tr>
		<tr>
			<td>G40 solution</td>
			<td>Serag Wiessner</td>
			<td>10755AAF</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelafundin ISO solution 40 mg/mL</td>
			<td>B. Braun</td>
			<td>210257641</td>
			<td></td>
		</tr>
		<tr>
			<td>Guidewire with marker</td>
			<td>Arrow</td>
			<td>14F21E0236</td>
			<td></td>
		</tr>
		<tr>
			<td>Haemostatic gauze (&#34;Tabotamp&#34;&#160; 5 x 7.5 cm)</td>
			<td>Ethicon</td>
			<td>474273</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin sodium 25,000IE</td>
			<td>Ratiopharm</td>
			<td>W08208A</td>
			<td></td>
		</tr>
		<tr>
			<td>Hico-Aquatherm 60</td>
			<td>Hospitalwerk</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Infusion Set Intrafix</td>
			<td>B.Braun</td>
			<td>4062981 L</td>
			<td></td>
		</tr>
		<tr>
			<td>Intrafix SafeSet 180 cm</td>
			<td>B.Braun</td>
			<td>4063000</td>
			<td></td>
		</tr>
		<tr>
			<td>Introcan Safety, 18 G&#160;</td>
			<td>B.Braun</td>
			<td>4251679-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluran CP</td>
			<td>CP-Pharma</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Large-bore venous catheter, 7Fr.</td>
			<td>Edwards Lifesciences</td>
			<td>I301F7</td>
			<td></td>
		</tr>
		<tr>
			<td>Ligaclip, medium</td>
			<td>Ethicon</td>
			<td>LT200</td>
			<td></td>
		</tr>
		<tr>
			<td>Ligaclip, small</td>
			<td>Ethicon&#160;</td>
			<td>LT100</td>
			<td></td>
		</tr>
		<tr>
			<td>Material scissors</td>
			<td>Martin&#160;</td>
			<td>11-285-23</td>
			<td></td>
		</tr>
		<tr>
			<td>Methylprednisolone (Urbason solubile forte 250 mg)</td>
			<td>Sanofi</td>
			<td>7823704</td>
			<td></td>
		</tr>
		<tr>
			<td>Monopolar ERBE ICC 300</td>
			<td>Fa. Erbe</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl solution (0.9%)</td>
			<td>Baxter</td>
			<td>1533</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Aesculap</td>
			<td>BM36</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Aesculap</td>
			<td>BM035R</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Aesculap</td>
			<td>BM 67</td>
			<td></td>
		</tr>
		<tr>
			<td>Neutral electrode</td>
			<td>Erbe Elektromedizin GmbH T&#252;bingen</td>
			<td>21191 - 060</td>
			<td></td>
		</tr>
		<tr>
			<td>Norepinephrine (Sinora)</td>
			<td>Sintetica GmbH</td>
			<td>04150124745717</td>
			<td></td>
		</tr>
		<tr>
			<td>Omniflush Sterile Filed 10 mL</td>
			<td>B.Braun</td>
			<td>3133335</td>
			<td></td>
		</tr>
		<tr>
			<td>Original Perfusorline 300 cm</td>
			<td>B.Braun</td>
			<td>21E26E8SM3</td>
			<td></td>
		</tr>
		<tr>
			<td>Overhold clamp</td>
			<td>INOX</td>
			<td>BH 959</td>
			<td></td>
		</tr>
		<tr>
			<td>Overhold clamp</td>
			<td>Ulrich</td>
			<td>CL 2911</td>
			<td></td>
		</tr>
		<tr>
			<td>Pentobarbital sodium(Release 500 mg/mL)</td>
			<td>WDT, Garbsen</td>
			<td>21217</td>
			<td></td>
		</tr>
		<tr>
			<td>Perfusers</td>
			<td>B.Braun</td>
			<td>49-020-031</td>
			<td></td>
		</tr>
		<tr>
			<td>Perfusor Syringe 50 mL</td>
			<td>B.Braun</td>
			<td>8728810F</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dishes&#160; 92 x 17 mm</td>
			<td>Nunc</td>
			<td>150350</td>
			<td></td>
		</tr>
		<tr>
			<td>Poole Suction Instrument Argyle flexibel</td>
			<td>Covidien, Mansfield USA</td>
			<td>20C150FHX</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium chloride (7.45%)</td>
			<td>B.Braun</td>
			<td>4030539078276</td>
			<td></td>
		</tr>
		<tr>
			<td>Pressure measurement set</td>
			<td>Codan pvb Medical GmbH</td>
			<td>957179</td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol (1%)</td>
			<td>CP-Pharma</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>S-Monovette 2.6 mL K3E</td>
			<td>Sarstedt</td>
			<td>04.1901</td>
			<td></td>
		</tr>
		<tr>
			<td>S-Monovette 2.9 mL 9NC</td>
			<td>Sarstedt</td>
			<td>04.1902</td>
			<td></td>
		</tr>
		<tr>
			<td>S-Monovette 7.5 mL Z-Gel</td>
			<td>Sarstedt</td>
			<td>11602</td>
			<td></td>
		</tr>
		<tr>
			<td>Sartinski clamp</td>
			<td>Aesculap</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel&#160; No.11</td>
			<td>Feather Safety Razor Co.LTD</td>
			<td>02.001.40.011</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>INOX&#160;</td>
			<td>BC 746</td>
			<td></td>
		</tr>
		<tr>
			<td>Seldinger Arterial catheter</td>
			<td>Arrow</td>
			<td>SAC-00520</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium bicarbonate (8.4%)</td>
			<td>B.Braun</td>
			<td>212768082</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterilization Set (&#34;ProSet Preparation Kit CVC&#34;)</td>
			<td>B.Braun</td>
			<td>4899719</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterofundin ISO solution</td>
			<td>B.Braun</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Suction</td>
			<td>Dahlhausen</td>
			<td>07.068.25.301</td>
			<td></td>
		</tr>
		<tr>
			<td>Suction Aesculap Securat 80</td>
			<td>Aesculap</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Suction catheter</td>
			<td>ConvaTec</td>
			<td>5365049</td>
			<td></td>
		</tr>
		<tr>
			<td>Sultamicillin (Unacid: 2000 mg Ampicillin/1000 mg Sulbactam)</td>
			<td>Pfizer</td>
			<td>DL253102</td>
			<td></td>
		</tr>
		<tr>
			<td>Suprapubic urinary catheter, &#34;bronchialis&#34;, 50 cm</td>
			<td>ConvaTec</td>
			<td>UK&#160; 1F02772</td>
			<td></td>
		</tr>
		<tr>
			<td>Suprasorb (&#34;Toptex lite RK&#34;)</td>
			<td>Lohmann &#38; Rauscher</td>
			<td>31654</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture Vicryl 3-0</td>
			<td>Ethicon</td>
			<td>VCP 1218 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture Vicryl 4-0</td>
			<td>Ethicon</td>
			<td>V392H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene 4-0</td>
			<td>Ethicon</td>
			<td>7588 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene 5-0, double armed</td>
			<td>Ethicon&#160;</td>
			<td>8890 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene 5-0, single armed</td>
			<td>Ethicon&#160;</td>
			<td>8720 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene 6-0, double armed</td>
			<td>Ethicon&#160;</td>
			<td>7230 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene 6-0, single armed</td>
			<td>Ethicon</td>
			<td>EH 7406 H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Prolene: blau 3-0&#160;</td>
			<td>Ethicon</td>
			<td>EH 7499H</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Safil 2/0</td>
			<td>Aesculap</td>
			<td>C 1038446</td>
			<td></td>
		</tr>
		<tr>
			<td>Suture, Terylene 0</td>
			<td>Serag Wiessner</td>
			<td>353784</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe 2 mL, 5 mL, 10 mL, 20 mL</td>
			<td>B.Braun</td>
			<td>4606027V</td>
			<td></td>
		</tr>
		<tr>
			<td>TransferSet &#34;1D/X-double&#34; steril 330 cm</td>
			<td>Fresenius Kabi AG</td>
			<td>2877101</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasound Butterfly IQ+</td>
			<td>Butterfly Network Inc.</td>
			<td>850-20014</td>
			<td></td>
		</tr>
		<tr>
			<td>Ventilator &#34;Oxylog Dr&#228;ger Fl&#34;</td>
			<td>Dr&#228;ger Medical AG</td>
			<td>No Catalog Number available</td>
			<td></td>
		</tr>
		<tr>
			<td>Yankauer Suction</td>
			<td>Medline</td>
			<td>RA19GMD</td>
			<td></td>
		</tr>
		<tr>
			<td>Zoletil 100 mg/mL&#160; (50 mg Zolazepam, 50 mg tiletamin)</td>
			<td>Virbac</td>
			<td>794-861794861</td>
			<td></td>
		</tr>
	</tbody>
</table>

porcine liver transplantation without veno-venous bypass as an extended criteria donor model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

The technique presented in this protocol has provided reliable and reproducible results in terms of hemodynamic stability and animal survival throughout the procedure, as well as graft function in the postoperative course.
Most recently, we applied the model for the study of ischemia-reperfusion injury and therapeutic interventions mitigating detrimental effects in the immediate postoperative course. Upon retrieval and 20 h of static cold storage, liver grafts (with a mean weight of 983.38 g) ...

Watch this Scientific Journal Video about Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model at JoVE.com

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...

Current issues in liver transplantation require the implementation of large animal models. This protocol describes the technique of porcine orthotopic liver transplantation without the use of a veno-venous bypass. This technique significantly reduces the logistical and technical complexity of the procedure.Furthermore, by engrafting livers after 20 hours of static cold storage, our model simulates extended criteria donor conditions. Begin by placing the anesthetized donor pig in a supine position and fix the limbs at the base of the operation table with elastic bands. Scrub the skin with an antiseptic agent and cover the animal with sterile drapes.After confirming the loss of petal reflex, perform a midline laparotomy beginning at the xiphoid process by using monopolar cautery and sever the falciform and the triangular ligaments using scissors and bipolar cautery. After sufficient liver dissection, incise the left portion of the diaphragm over a distance of 5 to 10 centimeters using scissors to locate the thoracic segment of the descending aorta. Encircle and place a 3-0 polyfilament ligature without tightening.Incise the right portion of the diaphragm over a distance of 5 to 10 centimeters using scissors and identify the suprahepatic vena cava inferior. Relocate the intestine to the upper left of the donor and enter the retroperitoneal space by transverse incision of the peritoneum over a distance of 5 to 10 centimeters using scissors. Place two 3-0 polyfilament ligatures around the abdominal aorta.One cranial of the iliac bifurcation and one approximately 3 centimeters cranially without tightening. Place another ligature around the infrahepatic vena cava inferior without tightening. Tighten the caudally-located first ligature around the abdominal aorta.After occluding the abdominal aorta cranially of the second ligature, make a transverse incision between both the ligatures using scissors. Insert the cannula into the incision and secure it with the remaining ligature. Using scissors, sever the suprahepatic inferior vena cava far cranially close to the right atrium.After blood loss of approximately 1, 500 to 2, 000 milliliters, cross-clamp the thoracic segment of the descending aorta by tying the ligature and start to antegrade perfusion. Tighten the ligature placed around the infrahepatic vena cava inferior. Incise the vessel cranially of the ligature and insert the surgical aspirator.After perfusion with 3, 500 milliliters of preservation solution over the course of approximately 10 to 15 minutes, sever the inside supra hepatic vena cava inferior of the donor pig, then sever the infrahepatic vena cava inferior at the level of the left renal vein. Sever the bile duct cranial of the pancreatic tissue between two 3-0 polyfilament ligatures to avoid bile spillage, then sever the portal vein cranial of the pancreas. Locate the celiac artery after blunt preparation and follow dorsally to the abdominal aorta.Excise the respective aortic segment to create a patch for the engraftment. Excise the diaphragm around the super hepatic vena cava inferior and sever the remaining adhesions using scissors. Extract the liver and place on ice.Perform a cholecystectomy or tighten a ligature around the cystic duct, then flush the common bile duct with at least 20 milliliters of preservation solution. Place the perfusion cannula into the portal vein and flush the graft with 500 milliliters of preservation solution. Place the graft in a sterile bowl placed on ice.Remove the lymphatic tissue of the aortic segment and the hepatic artery, thereby identifying and occluding the arterial side branches and lymphatic vessels with either clips, ligatures, or sutures. Proceed similarly with the portal vein. Perform shortening of the vessels and preparation of the aortic patch only upon and engraftment to take into account the individual anatomic circumstances.Identify the super hepatic vena cava inferior and place sutures around both diaphragmatic veins after removing surrounding diaphragmatic tissue. Place the anesthetized recipient pig in a supine position. Enter the abdominal cavity and sever the falciform and triangular ligaments of the anesthetized animal using scissors and bipolar cautery as demonstrated earlier.After sufficient liver dissection, encircle both the supra and infrahepatic vena cavae inferior close to the liver parenchyma. Incise the superficial peritoneal layer covering the hepatoduodenal ligament and identify the hepatic arteries before entering the liver parenchyma. Dissect using bipolar cautery or placing the clips, ligatures, or sutures.Dissect and sever the common bile duct below the cystic duct junction between two ligatures. Identify the portal vein and encircle close to the liver parenchyma. Identify the abdominal aorta after incising the avascular layer between the right and left diaphragmatic muscles.Prepare the aorta for aortic anastomosis by removal of the surrounding tissue. Perform recipient hepatectomy by placing in a traumatic vascular clamp on the portal vein and sever the portal vein close to the liver parenchyma followed by the infrahepatic vena cava. Place an atraumatic vascular clamp on the suprahepatic vena cava including the surrounding diaphragm while caudally retracting the liver and sever the vessel close to the liver parenchyma.Remove the recipient liver from the abdominal cavity. Place the donor liver into the abdominal cavity and shorten the donor or recipient suprahepatic vena cava inferior to an adequate length while avoiding kinking or too much tension on the anastomosis. Place a single 5-0 monofilament suture as a supporting thread adapting the right corner of the donor and recipient supra hepatic vena cavae inferior.Begin the dorsal side of the anastomosis from the left corner of the vessels with a 5-0 monofilament double-armed running suture. Remove the supporting thread when reaching the right corner and secure the running suture with a clamp, then continue with the ventral side of the anastomosis, again, beginning from the left corner of the vessels. After completing the ventral side of the anastomosis, tighten the suture with multiple knots without constricting the vessel diameter to avoid stenosis.Perform a vascular anastomosis of the donor and recipient portal vein as demonstrated earlier using a 6-0 monofilament double-armed suture. Begin the portal venous reperfusion by removing the vascular clamp, occluding the recipient portal vein and occlude the donor infrahepatic vena cava inferior with a vascular clamp after draining approximately 200 to 400 milliliters of blood. Slowly remove the vascular clamp occluding the recipient supra hepatic vena cava inferior and search for active bleeding.Shorten the donor and/or recipient infrahepatic vena cava inferior. then perform a vascular anastomosis of the donor and recipient infrahepatic vena cavae inferior. After clamping the abdominal aorta and making an incision to fit the patch, begin the aortic anastomosis with a 6-0 monofilament double-armed running suture at the cranial corner of the incision or patch.When reaching the caudal corner, secure the running suture with a clamp and complete the anastomosis again beginning at the cranial corner. Tighten the suture with multiple knots and slowly remove the vascular clamp after checking all vascular anastomoses. Place a catheter in the common bile duct and secure it with a single ligature.Close the abdomen temporarily by adapting the muscular fascia and the skin with a running suture and cover the abdomen with clinging film or drapes to avoid thermal loss. The mean operation time was 103.50 minutes including a mean and hepatic phase of 27.13 minutes. Only two recipients underwent an anhepatic phase of more than 30 minutes while all recipients showed declining lactate serum concentrations 4 hours after portal venous reperfusion.Recipients required increased concentrations of norepinephrine immediately before and throughout the anhepatic phase to stabilize the mean arterial pressure at 60 millimeters of mercury or higher. Low epinephrine concentrations were simultaneously used to increase cardiac output in this vulnerable time period. After engraftment, the mean arterial pressure and required doses of vasopressors remained stable.The LiMAx values obtained 6 hours after portal venous reperfusion were comparable to the values measured in the liver donors before organ procurement in all but one recipient with an anhepatic phase of 34 minutes. This model can be of special interest as a readout for workups focusing on mechanisms of ischemia and reperfusion injury and the reconditioning of extended criteria donor organs.

Research

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Medicine

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