<meta charset="utf8"></head><body>使用机器灌注的原位猪肝移植保存

<ol>
	<li>de Goeij, F., Schlegel, A., Muiesan, P., Guarrera, J. V., 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=Hypothermic+oxygenated+machine+perfusion+protects+from+cholangiopathy+in+donation+after+circulatory+death+liver+transplantation.">Hypothermic oxygenated machine perfusion protects from cholangiopathy in donation after circulatory death liver transplantation.</a> Hepatology. 74 (6), 3525-3528 (2021).</li><li>Dutkowski, P., 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=Evolving+trends+in+machine+perfusion+for+liver+transplantation.">Evolving trends in machine perfusion for liver transplantation.</a> Gastroenterology. 156 (6), 1542-1547 (2019).</li><li>Parente, A., 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+techniques+for+liver+transplantation:+A+meta-analysis+of+the+first+seven+randomized-controlled+trials.">Machine perfusion techniques for liver transplantation: A meta-analysis of the first seven randomized-controlled trials.</a> J Hepatol. 79 (5), 1201-1213 (2023).</li><li>Schlegel, A., 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=Outcomes+of+DCD+liver+transplantation+using+organs+treated+by+hypothermic+oxygenated+perfusion+before+implantation.">Outcomes of DCD liver transplantation using organs treated by hypothermic oxygenated perfusion before implantation.</a> J Hepatol. 70 (1), 50-57 (2019).</li><li>Zhang, Z. B., 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=Normothermic+machine+perfusion+protects+against+liver+ischemia-reperfusion+injury+during+reduced-size+liver+transplantation+in+pigs.">Normothermic machine perfusion protects against liver ischemia-reperfusion injury during reduced-size liver transplantation in pigs.</a> Ann Transplant. 24, 9-17 (2019).</li><li>Minor, 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=Hypothermic+reconditioning+by+gaseous+oxygen+improves+survival+after+liver+transplantation+in+the+pig.">Hypothermic reconditioning by gaseous oxygen improves survival after liver transplantation in the pig.</a> Am J Transplant. 11 (12), 2627-2634 (2011).</li><li>Linares-Cervantes, I., 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=Predictor+parameters+of+liver+viability+during+porcine+normothermic+ex+situ+liver+perfusion+in+a+model+of+liver+transplantation+with+marginal+grafts.">Predictor parameters of liver viability during porcine normothermic ex situ liver perfusion in a model of liver transplantation with marginal grafts.</a> Am J Transplant. 19 (11), 2991-3005 (2019).</li><li>Fu, 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=Porcine+partial+liver+transplantation+without+veno-venous+bypass:+an+effective+model+for+small-for-size+liver+graft+injury.">Porcine partial liver transplantation without veno-venous bypass: an effective model for small-for-size liver graft injury.</a> Transplant Proc. 43 (5), 1953-1961 (2011).</li><li>Brockmann, J. G., 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=Sequence+of+reperfusion+influences+ischemia/reperfusion+injury+and+primary+graft+function+following+porcine+liver+transplantation.">Sequence of reperfusion influences ischemia/reperfusion injury and primary graft function following porcine liver transplantation.</a> Liver Transpl. 11 (10), 1214-1222 (2005).</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> J Vis Exp. (99), e52055 (2015).</li><li>Ceresa, C., Nasralla, D., Pollok, J. M., Friend, P. 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> Nat Rev Gastroenterol Hepatol. 19 (3), 199-209 (2022).</li><li>Martins, P. N., Buchwald, J. E., Mergental, H., Vargas, L., Quintini, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+normothermic+machine+perfusion+in+liver+transplantation.">The role of normothermic machine perfusion in liver transplantation.</a> Int J Surg. 82, 52-60 (2020).</li><li>Br&#252;ggenwirth, I., 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=Prolonged+dual+hypothermic+oxygenated+machine+preservation+(DHOPE-PRO)+in+liver+transplantation:+Study+protocol+for+a+stage+2,+prospective,+dual-arm,+safety+and+feasibility+clinical+trial.">Prolonged dual hypothermic oxygenated machine preservation (DHOPE-PRO) in liver transplantation: Study protocol for a stage 2, prospective, dual-arm, safety and feasibility clinical trial.</a> BMJ Open Gastroenterol. 9 (1), 000842 (2022).</li><li>Mergental, 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=Transplantation+of+discarded+livers+following+viability+testing+with+normothermic+machine+perfusion.">Transplantation of discarded livers following viability testing with normothermic machine perfusion.</a> Nat Commun. 11 (1), 2939 (2020).</li><li>Nasralla, D., 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+randomized+trial+of+normothermic+preservation+in+liver+transplantation.">A randomized trial of normothermic preservation in liver transplantation.</a> Nature. 557 (7703), 50-56 (2018).</li><li>Melandro, 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=Viability+criteria+during+liver+ex-situ+normothermic+and+hypothermic+perfusion.">Viability criteria during liver ex-situ normothermic and hypothermic perfusion.</a> Medicina (Kaunas). 58 (10), 1434 (2022).</li><li>Warmuzi&#324;ska, N., &#321;uczykowski, K., Bojko, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+current+and+emerging+trends+in+donor+graft-quality+assessment+techniques.">Review of current and emerging trends in donor graft-quality assessment techniques.</a> J Clin Med. 11 (3), 487 (2022).</li><li>OuYang, Q., 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=Evaluation+of+the+ex+vivo+liver+viability+using+a+nuclear+magnetic+resonance+relaxation+time-based+assay+in+a+porcine+machine+perfusion+model.">Evaluation of the ex vivo liver viability using a nuclear magnetic resonance relaxation time-based assay in a porcine machine perfusion model.</a> Sci Rep. 11 (1), 4117 (2021).</li><li>Sampaziotis, 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=Cholangiocyte+organoids+can+repair+bile+ducts+after+transplantation+in+the+human+liver.">Cholangiocyte organoids can repair bile ducts after transplantation in the human liver.</a> Science. 371 (6531), 839-846 (2021).</li><li>van Leeuwen, O. B., 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=Transplantation+of+high-risk+donor+livers+after+ex+situ+resuscitation+and+assessment+using+combined+hypo-+and+normothermic+machine+perfusion:+A+prospective+clinical+trial.">Transplantation of high-risk donor livers after ex situ resuscitation and assessment using combined hypo- and normothermic machine perfusion: A prospective clinical trial.</a> Ann Surg. 270 (5), 906-914 (2019).</li><li>van Leeuwen, O. B., 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=Sequential+hypothermic+and+normothermic+machine+perfusion+enables+safe+transplantation+of+high-risk+donor+livers.">Sequential hypothermic and normothermic machine perfusion enables safe transplantation of high-risk donor livers.</a> Am J Transplant. 22 (6), 1658-1670 (2022).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anesthesia respirator</td>
			<td>Mindray,Shenzhen</td>
			<td>WATO EX-20&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic biochemical analyzer</td>
			<td>&#160;MNCHIP, China</td>
			<td>&#160;Celercare V5</td>
			<td></td>
		</tr>
		<tr>
			<td>Bama female miniature pigs&#160;</td>
			<td>Pearl Lab Animal Sci &#38; Tech Co,Ltd (Guangdong, China).&#160;</td>
			<td></td>
			<td>40-45 kg&#160;</td>
		</tr>
		<tr>
			<td>Blood gas analyzer&#160;</td>
			<td>&#160;Abbott</td>
			<td>i-STAT300</td>
			<td></td>
		</tr>
		<tr>
			<td>ECG monitor</td>
			<td>Shenzhen Ericon Medical Equipment Co., LTD China</td>
			<td>M-9000S</td>
			<td></td>
		</tr>
		<tr>
			<td>Fully automatic snowflake ice</td>
			<td>Changshu Shenghai Electric Co., Ltd. China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Perfusate in NMP</td>
			<td></td>
			<td></td>
			<td>5% Human serum albumin 100-150 mL, 
			Whole blood 1.2-1.5 L, 
			2.5% NaHCO3 21 mL, 
			10% CaCL2 7mL , 
			Heparin 5000 U , 
			Cefoxltin 1 g, 
			Metronidazole 500 mg, 
			Sodium taurocholate 5 g, 
			Short acting insulin 72 U, 
			Total parenteral nutrition solution 250-500 mL.</td>
		</tr>
		<tr>
			<td>Potal catheter</td>
			<td>Jinxin technology,Shunde,China</td>
			<td></td>
			<td>Portal vein catheter for custom cannula of varying internal diameter (6-8.5 mm)</td>
		</tr>
		<tr>
			<td>Refrigeration centrifuge&#160;</td>
			<td>hermo Fisher Scientific - CN</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>The CG8/CG4 blood gas test card</td>
			<td>&#160;Abbott</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>The CHEM 8 test card</td>
			<td>&#160;Abbott</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>The ex vivo liver machine perfuion device</td>
			<td>Devocean Medical Instrument Co., Ltd, Guangdong, China</td>
			<td>DEVOCEAN-LIVER 2000</td>
			<td>This is a multi-mode, temperature-controlled, biomimetic ex vivo liver machine perfusion device, capable of preserving the liver outside the body for 24 h</td>
		</tr>
		<tr>
			<td>UW Cold Storage solution&#160;&#160;</td>
			<td>Bridge to Life, Ltd., USA</td>
			<td>Belzer UW</td>
			<td>Liver in SCS group were preserved in UW Cold Storage solution&#160;</td>
		</tr>
		<tr>
			<td>UW Machine Perfusion Solution</td>
			<td>Bridge to Life, Ltd., USA</td>
			<td>Belzer MPS</td>
			<td>&#160;Adenine (free base) 0.68 g, 
			Calcium Chloride (dihydrate) 0.068 g, 
			Dextrose (+) 1.80 g, 
			Glutathione (reduced) 0.92 g, 
			HEPES (free acid) 2.38 g, 
			Hydroxyethyl Starch 50.0 g, 
			Magnesium Gluconate 1.13 g, 
			Mannitol 5.4 g, 
			Potassium Phosphate (monobasic) 3.4 g, 
			Ribose, D(-) 0.75 g, 
			Sodium Gluconate 17.45 g, 
			Sodium Hydroxide 0.70 g, 
			Sterile Water for Injection To 1000 mL Volume</td>
		</tr>
		<tr>
			<td>Vacuum extractor</td>
			<td>SMAF</td>
			<td>DYX-2A</td>
			<td></td>
		</tr>
	</tbody>
</table>

preservation of porcine donation after circulatory death (dcd) liver by perfusion and orthotopic liver transplantation

Liver transplantation remains the sole curative treatment for end-stage liver disease and selected liver cancers. Despite significant advancements in procurement, preservation, operative techniques, and post-transplant immunosuppression, a notable mortality rate persists among patients on the waiting list due to a shortage of suitable donor organs. A primary challenge lies in preserving livers procured from DCD, as these organs necessitate specialized care to mitigate ischemic injuries1. Ex vivo liver machine perfusion offers a unique method to both preserve and evaluate DCD liver grafts before transplantation2. Clinical trials have underscored the feasibility and safety of ex vivo liver machine perfusion for both standard and expanded criteria donors, employing either hypothermic or normothermic conditions3. Importantly, therapeutic interventions during ex vivo liver machine perfusion have shown promise in reducing ischemia-reperfusion injury (IRI)4.
In efforts to extend the preservation duration and enhance the quality of DCD liver grafts, ongoing animal experiments aim to optimize the performance of MP devices and refine the method of ex vivo liver preservation5. Porcine OLTx serves as an optimal model for clinically oriented research, validating the preservative quality of MP. However, ischemic injury to the donor, hemodynamic instability, and intestinal congestion during the anhepatic phase of porcine OLTx collectively impact the survival rate of the porcine model6,7.
A variable temperature-controlled MP device that integrates both NMP and DHOPE modes was utilized to preserve the DCD livers of porcine in the following protocol. This device facilitates extended ex vivo preservation of the DCD livers and alleviates the ischemic injury of the donor liver compared to traditional SCS. The apparatus ensures temperature regulation, supports long-distance transportation, and provides bionic perfusion alongside dynamic and accurate assessment of donor quality. The protocol contains all the information for a stable model of DCD liver preservation utilizing a sequential DHOPE-NMP mode followed by porcine OLTx, including the transition of perfusion settings, optimizing the anastomosis technique, and the procedure of the anhepatic phase.

<meta charset="utf8"></head><body>作者聚焦：肝移植机器灌注的进步和挑战

循环死亡 （DCD） 肝脏循环死亡 （DCD） 后通过灌注和原位肝移植保存猪捐献

Conventional static cold storage (SCS) exacerbates ischemic injury in the DCD liver, leading to severe complications for transplant recipients. To address ...

preservation-porcine-donation-after-circulatory-death-dcd-liver

Research

JoVE Journal

Bioengineering

Preservation of Porcine Donation after Circulatory Death (DCD) Liver by Perfusion and Orthotopic Liver Transplantation

222 次观看.  PLA General Hospital of Southern Theatre Command: People's Liberation Army General Hospital of Southern Theatre Command. 机器灌注技术促进了供体肝脏质量的保存，特别是对于扩大标准肝脏，但通过离心泵和氧合器持续泵送氧气和营养物质，它保持器官活动并监测肝功能，提高保存时间和运输成功率。使用机器灌注进行供体肝脏的主要挑战是用复杂的设备和熟练的操作来复制肝脏状况。站点后 DCD 肝脏模型的局限性包括肝脏质量、储存时间和肝脏期的恶化，导致实验?...

视频: 作者聚焦：肝移植机器灌注的进步和挑战

Conventional static cold storage (SCS) exacerbates ischemic injury in the DCD liver, leading to severe complications for transplant recipients. To address this issue, clinical application of MP technology for donor liver preservation is underway. Simultaneously, efforts are focused on the development of various MP instruments, validated through relevant animal model experiments. Effective large animal trials play a pivotal role in clinical applications. However, challenges persist in the ex vivo preservation of DCD livers and the transplantation procedure in pigs. These hurdles encompass addressing the prolonged preservation of donor livers, conducting viability tests, alleviating ischemic injuries, and shortening the anhepatic phase. The use of a variable temperature-controlled MP device facilitates the prolonged preservation of DCD livers through sequential Dual Hypothermic Oxygenated Machine Perfusion (DHOPE) and Normothermic Machine Perfusion (NMP) modes. This protocol enhances the porcine OLTx model by improving the quality of DCD livers, optimizing the anastomosis technique, and reducing the duration of the anhepatic phase.

This protocol outlines the methodology of establishing a porcine model utilizing variable temperature-controlled Machine Perfusion (MP) for the preservation of the donor liver, followed by orthotopic liver transplantation (OLTx). It aims to promote the success rate of OLTx using donor donation after circulatory death (DCD) liver and establish a stable model.