Name: ex situ normothermic machine perfusion of donor livers
Uploaded: 2015-05-26T15:00:00
Description: Watch this Scientific Journal Video about Ex Situ Normothermic Machine Perfusion of Donor Livers at JoVE.com

该研究工作得到由Innovatief Actieprogramma格罗宁根（IAG-3），扬德Kornelis屌斯蒂廷和提基亚Huizingafonds规定，所有在荷兰的资助。我们感谢所​​有荷兰移植协调员识别潜在的肝脏丢弃并取得知情同意。

该协议已获得医学伦理委员会的格罗宁根大学医学中心的（Medisch Ethische Toetsingscommissie），荷兰。 1.制备灌注流体注意：根据这个协议准备常温灌注机的灌注流体的总体积是2233毫升和灌注流体的目标容量渗透摩尔浓度是302 mOsmol /升。 <ol><li>从表1中描述的灌注流体的部件，保持人聚集的红血细胞，新鲜冰冻血浆和人白蛋白分离。混合各组分?...

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Hepatol. 58 (6), 1065-1067 (2013).</li><li>Gastaca, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biliary+complications+after+orthotopic+liver+transplantation:+A+review+of+incidence+and+risk+factors.">Biliary complications after orthotopic liver transplantation: A review of incidence and risk factors.</a> Transplant. Proc. 44 (6), 1545-1549 (2012).</li><li>Sanchez-Urdazpal, 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=Ischemic-type+biliary+complications+after+orthotopic+liver+transplantation.">Ischemic-type biliary complications after orthotopic liver transplantation.</a> Hepatology. 16 (1), 49-53 (1992).</li><li>Dubbeld, 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=Similar+liver+transplantation+survival+with+selected+cardiac+death+donors+and+brain+death+donors.">Similar liver transplantation survival with selected cardiac death donors and brain death donors.</a> Br. J. Surg. 97 (5), 744-753 (2010).</li><li>Guarrera, J. V., 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+machine+preservation+in+human+liver+transplantation:+The+first+clinical+series.">Hypothermic machine preservation in human liver transplantation: The first clinical series.</a> Am. J. Transplant. 10 (2), 372-381 (2010).</li><li>Henry, S. 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=Hypothermic+machine+preservation+reduces+molecular+markers+of+ischemia/reperfusion+injury+in+human+liver+transplantation.">Hypothermic machine preservation reduces molecular markers of ischemia/reperfusion injury in human liver transplantation.</a> Am. J. Transplant. 12 (9), 2477-2486 (2012).</li><li>Schlegel, A., Rougemont, O., Graf, R., Clavien, P. A., 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=Protective+mechanisms+of+end-ischemic+cold+machine+perfusion+in+DCD+liver+grafts.">Protective mechanisms of end-ischemic cold machine perfusion in DCD liver grafts.</a> J. Hepatol. 58 (2), 278-286 (2013).</li><li>op den Dries, S., 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+oxygenated+machine+perfusion+prevents+arteriolonecrosis+of+the+peribiliary+plexus+in+pig+livers+donated+after+circulatory+death.">Hypothermic oxygenated machine perfusion prevents arteriolonecrosis of the peribiliary plexus in pig livers donated after circulatory death.</a> PLoS. One. 9 (2), e88521 (2014).</li><li>Tolboom, 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=Subnormothermic+machine+perfusion+at+both+20+degrees+C+and+30+degrees+C+recovers+ischemic+rat+livers+for+successful+transplantation.">Subnormothermic machine perfusion at both 20 degrees C and 30 degrees C recovers ischemic rat livers for successful transplantation.</a> J. Surg. Res. 175 (1), 149-156 (2012).</li><li>Gringeri, E., 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=Subnormothermic+machine+perfusion+for+non-heart-beating+donor+liver+grafts+preservation+in+a+swine+model:+A+new+strategy+to+increase+the+donor+pool?+Transplant.">Subnormothermic machine perfusion for non-heart-beating donor liver grafts preservation in a swine model: A new strategy to increase the donor pool? Transplant.</a> Proc. 44 (7), 2026-2028 (2012).</li><li>Bruinsma, B. 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=Subnormothermic+machine+perfusion+for+ex+vivo+preservation+and+recovery+of+the+human+liver+for+transplantation.">Subnormothermic machine perfusion for ex vivo preservation and recovery of the human liver for transplantation.</a> Am. J. Transplant. 14 (6), 1400-1409 (2014).</li><li>op den Dries, S., 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=Ex+vivo+normothermic+machine+perfusion+and+viability+testing+of+discarded+human+donor+livers.">Ex vivo normothermic machine perfusion and viability testing of discarded human donor livers.</a> Am. J. Transplant. 13 (5), 1327-1335 (2013).</li><li>Sutton, M. E., 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=Criteria+for+viability+assessment+of+discarded+human+donor+livers+during+ex+vivo+normothermic+machine+perfusion.">Criteria for viability assessment of discarded human donor livers during ex vivo normothermic machine perfusion.</a> PLoS. One. 9 (11), (2014).</li><li>Minor, T., Efferz, P., Fox, M., Wohlschlaeger, J., Luer, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+oxygenated+rewarming+of+cold+stored+liver+grafts+by+thermally+graduated+machine+perfusion+prior+to+reperfusion.">Controlled oxygenated rewarming of cold stored liver grafts by thermally graduated machine perfusion prior to reperfusion.</a> Am. J. Transplant. 13 (6), 1450-1460 (2013).</li><li>Makowka, 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=Surgical+technique+of+orthotopic+liver+transplantation.">Surgical technique of orthotopic liver transplantation.</a> Gastroenterol. Clin. North. Am. 17 (1), 33-51 (1988).</li><li>Hansen, 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=Histological+examination+and+evaluation+of+donor+bile+ducts+received+during+orthotopic+liver+transplantation--a+morphological+clue+to+ischemic-type+biliary+lesion?.">Histological examination and evaluation of donor bile ducts received during orthotopic liver transplantation--a morphological clue to ischemic-type biliary lesion?.</a> Virchows Arch. 461 (1), 41-48 (2012).</li><li>Monbaliu, D., Brassil, 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:+Past,+present+and+future.">Machine perfusion of the liver: Past, present and future.</a> Curr. Opin. Organ. Transplant. 15 (2), 160-166 (2010).</li><li>Oldani, 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=Pre-retrieval+reperfusion+decreases+cancer+recurrence+after+rat+ischemic+liver+graft+transplantation.">Pre-retrieval reperfusion decreases cancer recurrence after rat ischemic liver graft transplantation.</a> J. Hepatol. 61 (2), 278-285 (2014).</li><li>Tromans, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature+and+pressure+dependent+solubility+of+oxygen+in+water:+A+thermodynamic+analysis.">Temperature and pressure dependent solubility of oxygen in water: A thermodynamic analysis.</a> Hydrometallurgy. 48 (3), 327-342 (1998).</li></ol>

此视频提供了使用设备，使压力控制双灌注通过肝动脉和门静脉一步的人供体肝脏的常温灌注机步协议。虽然按照此协议，灌注机的技术故障并没有出现，所有移植物灌注良好和良好的氧化。 易地肝脏灌注了血流动力学稳定，并为代谢活跃，所生产的胆16,17定义。 这是一个公认的协议，用于人类供体肝脏的灌注机。该技术具有超过SCS 21的常规方法的几?...

器官保存在肝移植目前的方法是冲洗出来，并与供肝的后续存储在寒冷的（0-4°C）保存液（如威斯康星溶液或组氨酸，色氨酸酮戊二酸溶液大学）。这种方法被称为静态冷库（SCS）。尽管肝脏在0-4℃下的代谢率是很低的，仍有0.27微摩尔氧/分钟/克肝组织，这是不能SCS 1期间提供的需求。 SCS的常规方法，因此，导致在一定程度的供体肝脏的（附加的）损伤。虽然这一数额保存损伤是不是质量好供肝的一个问题，它可以成为次优的肝脏已经遭受一定程度损伤的供体的一个关键和限制因素。出于这个原因，肝脏与次优的质量或所谓的扩展标准供体（ECD）肝脏经常拒绝用于移植的风险ö˚F早期移植失败被认为是太高了。对移植肾功能延迟，主要的非功能和非吻合口胆管狭窄（NAS）的高利率已经在肝脏接受者循环死亡（DCD），旧的捐助者或脂肪肝移植2收件人描述后从捐赠。 NAS是发病率和死亡率的肝脏移植后的一个主要原因。 NAS可能发生在两个预算外和捐助者的肝内胆管，可伴有导管内胆泥和铸造形成3,4。尽管NAS的病因被认为是多因素的，在移植保存和移植胆管缺血/再灌注损伤已被确定为主要的潜在机制2,5。一个DCD移植物的移植已被确定为最强的危险因素为NAS的发展中的一个。一段暖缺血的一个DCD供体，器官保存在冷缺血随后再灌注的组合，和在收件人损伤被认为是负责胆管可逆损伤，这在与胆管的不良再生能力结合，导致纤维化瘢痕和胆管的变窄肝移植2,5-之后。 NAS已经报道了高达接收DCD肝6-8 30％的患者。它已经很清楚，SCS的用于移植的当前方法肝移植是不足以preinjured ECD肝脏如来自DCD供体。需要其他方法来增加和优化利用ECD肝脏移植。 机灌注（MP）是器官保存，其可以提供供体器官的保存较好，相比SCS的方法。 MP可以是特别相关的ECD移植物的保存。 MP的一项重要优点是有可能向在保存期间提供氧气的移植物。 MP可以在各种温度下进行，已被列为低温（0-10℃），subnormothermic（10-36℃）和常温（36-37℃），MP（NMP）。取决于用于MP的温度，灌注流体的类型必须调整并随着温度的升高更多的氧气应提供。 MP在人肝移植的第一个临床应用是基于低温灌注未经灌注流体9,10的活性氧。在动物模型中，低温氧化的MP（0-10℃）已经显示出具有抗缺血的保护作用/肝的再灌注损伤移植物11，并提供更好的保存胆汁的胆管周围血管丛的管道12。 Subnormothermic含氧的MP在20℃或30℃下也进行了研究的动物模型，并且显示出提供早期恢复DCD肝脏移植物功能，相比SCS 13,14。人类肝脏subnormothermic充氧MP的可行性RECently报道了七人丢弃的供肝15。 NMP（37℃）允许接枝生存能力和现有功能的评估移植16,17。此外，MP允许肝移植物的逐渐复温移植前，这已被证明有利于恢复的移植物18和复苏。 在当前协议肝灌注机中使用的灌注装置能够双重灌注（通过门静脉和肝动脉）使用两个离心泵，即提供连续的门脉血流和脉动的动脉血流。该系统的压力进行控制，从而自动调节流经肝脏，取决于肝内阻力。两个中空纤维膜氧合允许肝移植物的氧合作用，以及用于除去的CO 2。该温度可根据期望的类型的MP（最小Temperat .. [温度设定<html> 10℃URE）。流量，压力和温度的显示装置实时地允许灌注过程的连续控制上。一种新的一次性无菌集油管，水库及氧合可用于每个移植（ 图1）的灌注。 这个视频文章的目的是提供一个一步一步的协议，使用这种新开发的肝脏灌注机人供体肝脏的易地常温灌注机。 <img alt="图1" src="/files/ftp_upload/52688/52688fig1.jpg" /> 图1：（A）的概略图，（B）中的灌注机的照片，（C）的人工肺的更近的视图，以及（D）用于人类供体肝脏的常温灌注离心泵。获得=“_空白”&gt;点击此处查看该图的放大版本。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Liver Assist</td>
			<td>Organ Assist</td>
			<td>OA.Li.Li.140</td>
			<td>Perfusion device</td>
		</tr>
		<tr>
			<td>Liver Assist disposable package</td>
			<td>Organ Assist</td>
			<td>OA.Li.DP.540</td>
			<td>Disposable set and cannulas</td>
		</tr>
		<tr>
			<td>Meredith No.8</td>
			<td>Vygon Nederlands B.V.</td>
			<td>1362082</td>
			<td>Bile duct cannula</td>
		</tr>
		<tr>
			<td>Human albumin 200g/l / ALBUMAN</td>
			<td>Sanquin</td>
			<td>15522598</td>
			<td>100 ml</td>
		</tr>
		<tr>
			<td>Modified parenteral nutrition</td>
			<td>Baxter Nederland B.V.</td>
			<td>N14G30E</td>
			<td>7.35 ml</td>
		</tr>
		<tr>
			<td>Multivitamins for infusion / CERNEVIT</td>
			<td>Baxter International Inc.</td>
			<td>9800927</td>
			<td>7 ul</td>
		</tr>
		<tr>
			<td>Concentrated trace elements for infusion / NUTRITRACE</td>
			<td>B. Braun Melsungen AG</td>
			<td>14811332</td>
			<td>7.35 ml</td>
		</tr>
		<tr>
			<td>Metronidazole 5mg/ml</td>
			<td>Baxter Nederland B.V.</td>
			<td>98181882</td>
			<td>40 ml</td>
		</tr>
		<tr>
			<td>Cefazoline / SERVAZOLIN</td>
			<td>Sandoz B.V.</td>
			<td>15611337</td>
			<td>2 ml</td>
		</tr>
		<tr>
			<td>Fast acting insulin&#160;</td>
			<td>various vendors</td>
			<td></td>
			<td>20 ml</td>
		</tr>
		<tr>
			<td>Calcium glubionate, intravenous solution 10%, 137.5 mg/mL</td>
			<td>Sandoz</td>
			<td>97038695</td>
			<td>40 ml</td>
		</tr>
		<tr>
			<td>Sterile H2O</td>
			<td>Fresenius Kabi Nederland B.V.</td>
			<td>98084453</td>
			<td>51.3 ml</td>
		</tr>
		<tr>
			<td>NaCl 0.9%</td>
			<td>Baxter Nederland B.V.</td>
			<td>15262510</td>
			<td>160 ml</td>
		</tr>
		<tr>
			<td>Heparin 5000 IE/ml for i.v. administration</td>
			<td>LEO Pharma B.V.</td>
			<td>98026178</td>
			<td>4 ml</td>
		</tr>
		<tr>
			<td>Sodium bicarbonate 8,4%</td>
			<td>B. Braun Melsungen AG</td>
			<td>97973874</td>
			<td>The amount depends on the pH</td>
		</tr>
		<tr>
			<td>Packed red blood cell (in SAGM)</td>
			<td>Blood bank (Sanquin)</td>
			<td>N0012000</td>
			<td>750 ml</td>
		</tr>
		<tr>
			<td>Fresh frozen plasma</td>
			<td>Blood bank (Sanquin)</td>
			<td>N04030A0/N04030B0</td>
			<td>900 ml</td>
		</tr>
	</tbody>
</table>

ex situ normothermic machine perfusion of donor livers

相对于传统的静态低温保存（0-4°C）， 易地灌注机可以提供更好的保护供肝。器官的连续灌流提供了机会，改善器官的质量，并允许移植前供肝易地可行性评估。这个视频文章提供了分步协议易地人供体肝脏的常温灌注机（37°C）使用的设备，提供了肝动脉和门静脉的连续灌流的压力和温度控制脉冲灌流。灌注流体由两个中空纤维膜氧合充氧，温度可以在10°C和37℃下进行调节。在灌注期间，肝脏的代谢活性以及损伤的程度可以通过从灌注流体取样的生化分析进行评估。灌注机是一个非常有前途的工具增加肝脏适合于移植的数量。

这是由于各种原因拒绝移植12人的肝脏获得知情同意从捐赠者家庭的调研后使用。捐赠者的特征是通过使用本文中描述的协议表2中描述的人类供体肝脏灌流normothermically 6小时。肝移植物的质量被监测肝灌注（ - D图2A）的宏观均匀性进行评价。肝脏的血流动力学通过监测变化的动脉和门的流动进行评估。在肝动脉和门静脉流和流的随后稳定的初始增加进行观察，结果为256±1...

Watch this Scientific Journal Video about Ex Situ Normothermic Machine Perfusion of Donor Livers at JoVE.com

Ex Situ Normothermic Machine Perfusion of Donor Livers

在这里，我们提出描述体肝移植含氧易地灌注机的协议。本文包含一步步协议采购和准备肝移植的灌注机，准备灌注液，素灌注机，并执行肝移植的含氧常温灌注机。

供肝易地常温灌注机

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

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Ex Situ Normothermic Machine Perfusion of Donor Livers

Single Port Donor Nephrectomy

单端口捐赠者肾切除术

In 2007, Rane presented the first single port nephrectomy for a small non-functioning kidney at the World Congress of Endourology.  Since that time, the ...

科研

医学

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

Subnormothermic的技术<em&gt;前体内</em&gt;肝脏灌注的存储，评估和修复边际移植肝

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for ...

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

步骤人类隐静脉<em&gt;前体内</em&gt;灌注和外部强化

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion

功能人类肝脏保存和恢复由Subnormothermic机灌注手段

There is currently a severe shortage of liver grafts available for transplantation.  Novel organ preservation techniques are needed to expand the pool of ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

常温<em&gt;离体</em&gt;肾脏的灌注肾的移植保护移植前

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

体外常温肝灌注的小动物模型

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Normothermic Ex Situ Heart Perfusion in Working Mode: Assessment of Cardiac Function and Metabolism

工作模式下的心功能和代谢评估

The current standard method for organ preservation (cold storage, CS), exposes the heart to a period of cold ischemia that limits the safe preservation ...

Establishing In Situ Closed Circuit Perfusion of Lower Abdominal Organs and Hind Limbs in Mice

在小鼠中建立下腹部器官和后肢的原点闭路灌注

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small ...

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

非原位负压通气肺灌注：肺功能和代谢的评估

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

A Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

一种猪异位心脏移植方案，用于向心脏同种异体移植物提供治疗药物

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...