Name: normothermic ex situ heart perfusion in working mode: assessment of cardiac function and metabolism
Uploaded: 2019-01-12T12:30:00
Description: Watch this Scientific Journal Video about Normothermic Ex Situ Heart Perfusion in Working Mode: Assessment of Cardiac Function and Metabolism at JoVE.com

这项工作得到了加拿大国家移植研究方案赠款的支持。sh 是一个医学和牙科学院的学生在心脏科学的研究生。dhf 接受了国家科学和工程研究委员会和加拿大卫生研究所的合作研究项目 (chrp) 赠款。

本手稿中的所有程序都是按照加拿大动物护理理事会的指导原则和实验室动物护理和使用指南进行的。这些议定书得到了艾伯塔大学动物护理机构委员会的批准。该方案已应用于幼小约克郡猪35-50 公斤之间。所有参与 eshp 程序的个人都接受了适当的生物安全培训。
1. 手术前准备
<ol><li>将风琴室正确地放置在仪器车上, 并将硅支撑膜安装在风琴室内。在图 1</str...

<ol>
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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preserving+and+evaluating+hearts+with+ex+vivo+machine+perfusion:+An+avenue+to+improve+early+graft+performance+and+expand+the+donor+pool.">Preserving and evaluating hearts with ex vivo machine perfusion: An avenue to improve early graft performance and expand the donor pool.</a> European Journal of Cardiothoracic Surgery. 34 (2), 318-325 (2008).</li><li>Freed, D. H., White, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor+heart+preservation:+Straight+up,+or+on+the+rocks?.">Donor heart preservation: Straight up, or on the rocks?.</a> Lancet. 385 (9987), 2552-2554 (2015).</li><li>Guibert, E. 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=Organ+preservation:+Current+concepts+and+new+strategies+for+the+next+decade.">Organ preservation: Current concepts and new strategies for the next decade.</a> Transfusion Medicine and Hemotherapy. 38 (2), 125-142 (2011).</li><li>Collins, M. 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=Use+of+diffusion+tensor+imaging+to+predict+myocardial+viability+after+warm+global+ischemia:+Possible+avenue+for+use+of+non-beating+donor+hearts.">Use of diffusion tensor imaging to predict myocardial viability after warm global ischemia: Possible avenue for use of non-beating donor hearts.</a> Journal of Heart and Lung Transplantation. 26 (4), 376-383 (2007).</li><li>White, C. W., 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+cardioprotective+preservation+strategy+employing+ex+vivo+heart+perfusion+facilitates+successful+transplant+of+donor+hearts+after+cardiocirculatory+death.">A cardioprotective preservation strategy employing ex vivo heart perfusion facilitates successful transplant of donor hearts after cardiocirculatory death.</a> Journal of Heart and Lung Transplantation. 32 (7), 734-743 (2013).</li><li>Iyer, 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=Normothermic+ex+vivo+perfusion+provides+superior+organ+preservation+and+enables+viability+assessment+of+hearts+from+DCD+donors.">Normothermic ex vivo perfusion provides superior organ preservation and enables viability assessment of hearts from DCD donors.</a> American Journal of Transplantation. 15 (2), 371-380 (2015).</li><li>Peltz, 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=Perfusion+preservation+maintains+myocardial+ATP+levels+and+reduces+apoptosis+in+an+ex+vivo+rat+heart+transplantation+model.">Perfusion preservation maintains myocardial ATP levels and reduces apoptosis in an ex vivo rat heart transplantation model.</a> Surgery. 138 (4), 795-805 (2005).</li><li>Liao, R., Podesser, B. K., Lim, C. 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+continuing+evolution+of+the+Langendorff+and+ejecting+murine+heart:+New+advances+in+cardiac+phenotyping.">The continuing evolution of the Langendorff and ejecting murine heart: New advances in cardiac phenotyping.</a> American Journal of Physiology-Heart and Circulatory Physiology. 303 (2), H156-H167 (2012).</li><li>Rivard, L., Gallegos, R., Ogden, I., Bianco, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perfusion+Preservation+of+the+Donor+Heart:+Basic+Science+to+Pre-Clinical.">Perfusion Preservation of the Donor Heart: Basic Science to Pre-Clinical.</a> Journal of Extra Corporeal Technology. 41 (3), 140-148 (2009).</li><li>Dhital, K. K., 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=Adult+heart+transplantation+with+distant+procurement+and+ex+vivo+preservation+of+donor+hearts+after+circulatory+death:+A+case+series.">Adult heart transplantation with distant procurement and ex vivo preservation of donor hearts after circulatory death: A case series.</a> Lancet. 385 (9987), 2585-2591 (2015).</li><li>Messer, S., Ardehali, A., Tsui, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normothermic+donor+heart+perfusion:+Current+clinical+experience+and+the+future.">Normothermic donor heart perfusion: Current clinical experience and the future.</a> Transplant International. 28 (6), 634-642 (2015).</li><li>White, C. W., 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=Assessment+of+donor+heart+viability+during+ex+vivo+heart+perfusion.">Assessment of donor heart viability during ex vivo heart perfusion.</a> Canadian Journal of Physiology and Pharmacology. 93 (10), 893-901 (2015).</li><li>Messer, S. 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=Functional+assessment+and+transplantation+of+the+donor+heart+after+circulatory+death.">Functional assessment and transplantation of the donor heart after circulatory death.</a> Journal of Heart and Lung Transplantation. 35 (12), 1443-1452 (2016).</li><li>Hatami, 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=Endoplasmic+reticulum+stress+in+ex+vivo+heart+prfusion:+A+comparison+between+working+vs+non-working+modes.">Endoplasmic reticulum stress in ex vivo heart prfusion: A comparison between working vs non-working modes.</a> Canadian Journal of cardiology. 33 (10), (2017).</li><li>White, C. W., 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+perfusion+in+a+loaded+state+improves+the+preservation+of+donor+heart+function.">Ex vivo perfusion in a loaded state improves the preservation of donor heart function.</a> Canadian Journal of cardiology. 31 (10), s202 (2015).</li><li>White, C. W., 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+wholeblood-based+perfusate+provides+superior+preservation+of+myocardial+function+during+ex+vivo+heart+perfusion.">A wholeblood-based perfusate provides superior preservation of myocardial function during ex vivo heart perfusion.</a> Journal of Heart and Lung Transplantation. (14), (2014).</li><li>Lips, D. 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=Left+ventricular+pressure-volume+measurements+in+mice:+comparison+of+closed-chest+versus+open-chest+approach.">Left ventricular pressure-volume measurements in mice: comparison of closed-chest versus open-chest approach.</a> Basic Research in Cardiology. 99 (5), 351-359 (2004).</li><li>Morita, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+there+a+crystal+ball+for+predicting+the+outcome+of+cardiomyopathy+surgery?+Preload+recruitable+stroke+work,+may+be+a+possible+candidate.">Is there a crystal ball for predicting the outcome of cardiomyopathy surgery? Preload recruitable stroke work, may be a possible candidate.</a> Journal of Cardiology. 71 (4), 325-326 (2018).</li><li>Hatami, 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=.">.</a> Canadian Society for Transplantation. , (2017).</li><li>Anthony, 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=Ex+vivo+coronary+angiographic+evaluation+of+a+beating+donor+heart.">Ex vivo coronary angiographic evaluation of a beating donor heart.</a> Circulation. 130 (25), e341-e343 (2014).</li><li>Sandha, J. K., 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=Steroids+Limit+Myocardial+Edema+During+Ex+vivo+Perfusion+Of+Hearts+Donated+After+Circulatory+Death.">Steroids Limit Myocardial Edema During Ex vivo Perfusion Of Hearts Donated After Circulatory Death.</a> Annals of Thoracic Surgery. , (2018).</li></ol>

成功灌注是根据研究的目的来定义的;但是, 这应该包括不间断的 eshp 所需的时间量和完整的收集有关心功能的数据在灌注过程中。为此, 必须遵循议定书中的几个关键步骤。
心脏是一个对氧气和能量有很高需求的器官, 最大限度地减少插管和灌注前的缺血时间是必须遵循的重要原则。采购过程、将心脏安装在 eshp 设备上以及启动灌注不应超过20-30分钟。
为...

临床相关性
虽然自1967年第一次心脏移植以来, 心脏移植的大多数方面都有了显著的发展, 但冷藏 (cs) 仍然是供体心脏保护的标准 1。cs 使器官暴露在冷缺血期, 限制了安全保存间隔 (4-6小时), 并增加了原发性移植物功能障碍2,3,4的风险。由于 cs 的静态性质, 在器官采购和移植之间的时间内, 无法评估功能或治疗干预。这是扩大标准捐助方的一个特别限制, 包括循环死亡后捐赠的心脏, 这给克服需求与目前捐助方之间的巨大差距造成了障碍 5、6.为了解决这一局限性, 提出了一种新的半生理方法来保存捐赠的心脏, 通过在保存时向心脏提供含氧、营养丰富的香水, 最大限度地减少冷缺血的暴露1,7.,8。
原位心脏灌注
在孤立的心脏的异地检查中最常用的方法之一是朗根多夫灌注。在1895年由 oskar langendorff 介绍的这种方法中, 血液流入冠状动脉, 流出孤立心脏的冠状窦, 心脏处于空的和跳动的状态9,10.使用转移医疗器官护理系统设备 (ocs) 在 langendorff 模式下的临床 eshp 已被证明在保存标准供体心脏1方面不低于 cs, 并为 dcd 心脏的临床移植提供了便利11. 然而, 人们对该装置评价器官可行性的能力表示关切, 因为一些最初被认为可移植的捐献者心脏在 ocs3上灌注后被丢弃。ocs 在 langendorff (非工作) 模式下支持心脏, 因此具有有限的能力来评估心脏的抽水功能3,12。越来越多的证据表明, 功能参数为评估器官生存能力提供了更好的方法, 这表明在 eshp 3 期间, 评估心脏功能可能成为评估和选择移植心脏的可靠工具 此外,我们对原位灌注猪心脏的研究表明, 在工作模式下, eshp 为心脏提供了增强的功能保护。灌注间隔15,16。
能够在工作模式下保持心脏的 eshp 设备必须具有一定的自动化级别, 以安全、精确地保持预紧力、后负荷和流量。此外, 这种系统应具有灵活性, 以促进对心脏功能进行全面评估。这里使用的 eshp 设备配备了定制软件, 1) 提供和维护所需的主动脉 (o) 和左心房 (la) 压力流和 2) 提供功能参数的实时分析和压力波形的视觉评估。对监督的最低需求。利用标准充液压力传感器采集压力数据, 利用跨时间多普勒流量探头采集流量数据。这些信号分别通过桥接和模拟输入进行数字化。心脏水平定位, 轻微提升到柔软的硅胶膜上的大血管。插管附件穿过膜, 加入一个顺应性室, 用于抑制心室射血。这项工作的目的是为心脏移植领域的研究人员提供一个协议, 用于对心脏进行原位灌注和评估, 在正常的, 半生理条件下的工作模式, 在一个大型哺乳动物 (约克郡猪) 模型。

<table>
	<tbody>
		<tr>
			<td>Debakey-Metzenbaum dissecting scissors</td>
			<td>Pilling</td>
			<td>342202</td>
			<td></td>
		</tr>
		<tr>
			<td>MAYO dissecting scissors</td>
			<td>Pilling</td>
			<td>460420</td>
			<td></td>
		</tr>
		<tr>
			<td>THUMB forceps</td>
			<td>Pilling</td>
			<td>465165</td>
			<td></td>
		</tr>
		<tr>
			<td>Debakey straight vascular tissue forceps&#160;</td>
			<td>Pilling</td>
			<td>351808</td>
			<td></td>
		</tr>
		<tr>
			<td>CUSHING Gutschdressing forceps</td>
			<td>Pilling</td>
			<td>466200</td>
			<td></td>
		</tr>
		<tr>
			<td>JOHNSON needle holder</td>
			<td>Pilling</td>
			<td>510312</td>
			<td></td>
		</tr>
		<tr>
			<td>DERF needle holder</td>
			<td>Pilling</td>
			<td>443120</td>
			<td></td>
		</tr>
		<tr>
			<td>Sternal saw</td>
			<td>Stryker</td>
			<td>6207</td>
			<td></td>
		</tr>
		<tr>
			<td>Sternal retractor</td>
			<td>Pilling</td>
			<td>341162</td>
			<td></td>
		</tr>
		<tr>
			<td>Vorse tubing clamp</td>
			<td>Pilling</td>
			<td>351377</td>
			<td></td>
		</tr>
		<tr>
			<td>MORRIS ascending aorta clamp</td>
			<td>Pilling</td>
			<td>353617</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical snare (tourniquet) set</td>
			<td>Medtronic</td>
			<td>CVR79013</td>
			<td></td>
		</tr>
		<tr>
			<td>2-0 SILK black 12 X 18&#34; strands</td>
			<td>ETHICON</td>
			<td>A185H</td>
			<td></td>
		</tr>
		<tr>
			<td>3-0 PROLENE blue 18&#34; PS-2 cutting</td>
			<td>ETHICON</td>
			<td>8687H</td>
			<td></td>
		</tr>
		<tr>
			<td>Biomedicus pump drive (modified)</td>
			<td>Medtronic</td>
			<td>540</td>
			<td>Modified to allow remote electronic control of pump speed</td>
		</tr>
		<tr>
			<td>Biomedicus pump</td>
			<td>Maquet</td>
			<td>BPX-80</td>
			<td></td>
		</tr>
		<tr>
			<td>Membrane oxigenator D 905</td>
			<td>SORIN GROUP</td>
			<td>50513</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubing flow module&#160;&#160;</td>
			<td>Transonic</td>
			<td>Ts410</td>
			<td></td>
		</tr>
		<tr>
			<td>PXL clamp-on flow sensor</td>
			<td>Transonic</td>
			<td>ME9PXL-BL37SF</td>
			<td></td>
		</tr>
		<tr>
			<td>TruWave pressure transducer</td>
			<td>Edwards</td>
			<td>VSYPX272</td>
			<td></td>
		</tr>
		<tr>
			<td>Intercept tubing 3/8&#34; X 3/32&#34; X 6&#39;</td>
			<td>Medtronic</td>
			<td>3506</td>
			<td></td>
		</tr>
		<tr>
			<td>Intercept tubing 1/4&#34; X 1/16&#34; X 8&#39;</td>
			<td>Medtronic</td>
			<td>3108</td>
			<td></td>
		</tr>
		<tr>
			<td>Heated/Refrigerated Bath Circulator&#160;</td>
			<td>Grant</td>
			<td>TX-150</td>
			<td></td>
		</tr>
		<tr>
			<td>ABL 800 FLEX Blood Gas Analyzer</td>
			<td>Radiometer</td>
			<td>989-963</td>
			<td></td>
		</tr>
		<tr>
			<td>5F Ventriculr straight pigtail cathter</td>
			<td>CORDIS</td>
			<td>534550S</td>
			<td></td>
		</tr>
		<tr>
			<td>5F AVANTI+ Sheath Introducer</td>
			<td>CORDIS</td>
			<td>504605A</td>
			<td></td>
		</tr>
		<tr>
			<td>Emerald Amplatz Guidewire</td>
			<td>CORDIS</td>
			<td>502571A</td>
			<td></td>
		</tr>
		<tr>
			<td>Dual chamber pace maker</td>
			<td>Medtronic</td>
			<td>5388</td>
			<td></td>
		</tr>
		<tr>
			<td>Defibrilltor</td>
			<td>CodeMaster</td>
			<td>M1722B</td>
			<td></td>
		</tr>
		<tr>
			<td>Infusion pump</td>
			<td>Baxter</td>
			<td>AS50</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical electrocautery device</td>
			<td>Kls Martin</td>
			<td>ME411</td>
			<td></td>
		</tr>
		<tr>
			<td>Gas mixer</td>
			<td>SECHRIST</td>
			<td>3500 CP-G</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical oxygen tank</td>
			<td>praxair</td>
			<td>2014408</td>
			<td></td>
		</tr>
		<tr>
			<td>Cabon dioxide tank</td>
			<td>praxair</td>
			<td>5823115</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td>MP biomedicals</td>
			<td>218057791</td>
			<td></td>
		</tr>
	</tbody>
</table>

normothermic ex situ heart perfusion in working mode: assessment of cardiac function and metabolism

目前的标准器官保存方法 (冷库, cs), 使心脏暴露在冷缺血期, 限制了安全保存时间, 增加了移植后不良结局的风险。此外, cs 的静态性质不允许在保存间隔内进行器官评估或干预。诺曼原位心脏灌注 (eshp) 是一种新的方法, 保存捐赠的心脏, 通过提供含氧, 营养丰富的心脏吸乳, 最大限度地减少冷缺血。eshp 在保存标准供体心脏方面已被证明不低于 cs, 也为循环死亡测定后捐献的心脏的临床移植提供了便利。目前, 唯一可用的临床 eshp 设备在卸载、非工作状态下灌注心脏, 限制了对心肌性能的评估。相反, 在工作模式下的 eshp 提供了机会, 通过评估生理条件下的功能和代谢参数, 全面评估心脏表现。此外, 早期的实验研究表明, eshp 在工作模式下可能会导致更好的功能保存。在这里, 我们描述了在一个大型哺乳动物 (猪) 模型中对心脏进行原位灌注的协议, 该模型可针对不同的动物模型和心脏大小进行重现。此 eshp 设备中的软件程序允许实时和自动控制泵的速度, 以保持所需的主动脉和左心房压力, 并评估各种功能和电生理参数, 而对监督/操纵。

在灌注开始时 (在非工作模式下), 当系统和灌注的温度接近正常时, 心脏通常会恢复窦性心律。当进入工作模式时, 当 la 压力接近所需的值时, 应观察到对 o 压力跟踪的喷射, 并应逐渐增加 la 流量 (心脏输出量的反射)。在约克郡猪模型 (35-50 公斤) 和启动心脏重量180-220 克, 最初的 la 流量将是约 2, 000 mL/min, 这通常将接近约 2, 750 mL/min 在工作模式灌注的第一个小时。<strong class="...

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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

诺本心脏原位灌注 (eshp), 使心脏保持跳动, 半生理状态。在工作模式下进行操作时, eshp 提供了对捐献者心脏功能和器官生存能力进行复杂评估的机会。在这里, 我们描述了我们的方法在 eshp 期间心肌性能评估。

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

该方法有助于回答有关捐赠者心脏的最佳保存以及其功能和代谢评估的后续移植适宜性的关键问题。这种技术的主要优点是，它允许对供体心脏进行功能评估，而这种评估并不完全基于间接代谢评估。这种技术的影响延伸到功能失调的捐赠者心脏的治疗，因为心脏功能可以很容易地和反复评估随着时间的推移。一般来说，对这件事的技术人员将很难做，因为以安全可靠的方式连接心脏需要一些技术专长。此方法的可视化演示至关重要，因为心脏检索和连接到设备的技术方面可能很难仅通过文本教学进行学习。在确认对有毒刺激缺乏反应后，对麻醉猪进行中位胸切除术，一旦心脏暴露，使用 Metzenbaum 剪刀打开胸腺。使用1-0丝线固定胸骨的心缘，并逐渐从两阶段静脉管中收集750毫升全血，放在自动 <3> <3>化玻璃容器的右中庭，在15分钟内，同时用一升同位素晶体溶液取代体积。将血液加入灌注回路，用750毫升的Krebs-Henseleit缓冲液补充8%白蛋白，并将14个量表心肺痛针和管管放入上升大道。用圈套将心痛针固定在大血管上，将心痛管连接到心痛包。在400毫升心肺痛中加入100毫升的血液。开气当结束之后，用大音夹交叉夹住上升的大音管。接下来，将心肺解送到心大音根。当所有溶液都施用后，拆下夹子并切除心脏，包括所有大音管和下降大音的一段。为了防止长期缺血的有害影响，心脏应尽快采购并安装在仪器上进行灌注。去除左心房周围多余的组织后，使用 3-0 聚丙烯缝合线在左心房孔周围放置一个钱包串缝合线。用 3 - 0 聚丙烯缝合并关闭劣质的维纳卡瓦。将左心房管放入左心房孔中，用圈套固定该角膜。将左心房管放在仪器的硅胶膜嵌入磁铁上，注意左心房的磁铁和相应的金属环正确接合。将大音管连接到硅胶膜中嵌入的大音管上，并使用丝绸领带将大音管固定在管周围。转动大音管，实现适当的谎言，没有张力或扭结，将大奥泵速度提高至约 1600 RPM，轻轻挤压心室以脱气心脏。大音根中剩余的空气将穿过无名和亚克拉维安分支。将主动脉净化线连接到非主动脉，用丝绸领带固定连接。用丝绸领带捕捉左亚克拉维亚动脉孔，用圈套和扣子固定闭口。通过亚冠动脉的孔放置一个5个法国导管护套，调整导管的长度和方向，使导管不干扰主动脉瓣功能。调整大音泵速度，达到 30 毫米汞的均值压力，以在非工作兰根多夫模式下开始灌注。肺动脉线中暗脱氧的过氧甲酸酯的出现表明冠状动脉流的重建。将热交换器调至 38 摄氏度以加热香水，并使用血气分析仪检查溶解气体状态，将气体混合物调整为适当的参数。在非工作模式下一小时后，在软件所需的左心房压力部分输入合适的左心房压力，并启动反馈回路。激活的工作模式将显示为绿色按钮，左心房泵速度将自动增加和降低，以达到并保持所选的左心房压力。为了评估和记录稳态数据，请将充满液体的辫子导管通过引入器护套放入左心室。为了评估预装可招募冲程工作，请从左心室取出辫子导管，并在分析期间调整左心房泵速度的所需下降速率，以及分析的所需时间段。单击记录压力音量循环。该软件将自动退出工作模式，并逐渐降低左心房泵转速，同时记录左心室冲程工作和左心房压力。在数据收集结束时，软件将执行新获取的数据集的线性回归，以生成预加载可招募冲程工作。原位器心脏软件完成分析后，主页上将出现一条消息，显示分析的相关性系数。如果系数大于 0.95，请单击"确定"。将记录预加载可招募的中风工作分析结果。要评估心脏和呼吸液的代谢状态，请使用从从动脉和肺动脉线采集的呼吸道样本的血气分析中获得的信息，每一到两个小时。在起始心脏重量为180至220克的约克郡猪模型中，最初的左心房流量约为2000毫升分钟，通常在工作模式的第一到两个小时的灌注中接近每分钟约2，250毫升。对在原位心脏灌注过程中获得的富酸盐样本进行的血气分析和代谢评估提供了有关心脏代谢状态的大量信息，表明富酸盐乳酸盐浓度和计算心肌氧气消耗所需的信息。在没有自然代谢其清晰心脏生物标志物的器官的情况下，通常观察到这些因子在香水溶液中的积累。除了负载相关参数外，左心室预载可招募冲程工作还可以在计算机控制的线性降低左心房压力期间进行评估，如所证明的。原位心脏灌注期间的超声心动图也可以提供有关心肌功能和解剖参数的其他信息。在尝试此程序时，重要的是要记住，原位心脏灌注仍处于起步阶段，需要进行更多的研究，以设计更好的代谢心脏支持，以更好地保持心脏活力和功能。按照这个程序，其他方法，如血管成像可以执行，以回答有关捐赠者心脏的冠状动脉状态的其他问题。这项技术开发后，为器官移植领域的研究人员探索在患者等待心脏移植时安全移植延长标准捐赠心脏的可能性铺平了道路。不要忘记，使用生物材料可能非常危险，在执行此过程时应始终采取预防措施，如戴手套和防护眼镜。

Research

JoVE Journal

Medicine

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

使用小动物看更衣室反转恢复（SALLI）的MRI大鼠心脏功能和心肌形态学评估

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

科研

医学

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型：教训制定鼠EVLP课程中学

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

Ex Situ Normothermic Machine Perfusion of Donor Livers

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 ...

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, ...

Echocardiographic Assessment of Cardiac Anatomy and Function in Adult Rats

成人大鼠心脏解剖与功能的超声心动图评估

The use of experimental animal models has become crucial in cardiovascular science. Most studies using rodent models are focused on two-dimensional ...

In Vitro Assessment of Cardiac Function Using Skinned Cardiomyocytes

使用皮肤心肌细胞进行心脏功能的体外评估

In this article, we describe the steps required to isolate a single permeabilized (&#34;skinned&#34;) cardiomyocyte and attach it to a force-measuring ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

诱导大鼠心肌梗死的半微创方法及独立工作心脏系统对心脏功能的评估

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...

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 ...

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

非原生离体胰腺灌注用于移植前胰腺同种异 体 移植物的保存

Pancreas transplantation (PTx) is a curative treatment for people who live with the burden of a diagnosis of diabetes mellitus (DM). However, due to organ ...