Name: left lung orthotopic transplantation in a juvenile porcine model for eslp
Uploaded: 2022-02-14T13:30:00
Description: Watch this Scientific Journal Video about Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP at JoVE.com

这项研究由大学医院基金会资助。

所有程序均按照加拿大动物护理委员会的指导方针和实验动物的护理和使用指南进行。这些协议得到了阿尔伯塔大学机构动物护理委员会的批准。该方案已应用于35-50公斤的雌性幼年约克郡猪。猪是无病原体的食品级标本。它们是从加拿大艾伯塔省埃德蒙顿的猪研究和技术中心购买的（https://srtc.ualberta.ca）。所有参与ESLP程序的人都接受了适当的生物安全培训。
1.术前准备和麻?...

<ol>
	<li>Chambers, D. 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=The+international+thoracic+organ+transplant+registry+of+the+international+society+for+heart+and+lung+transplantation:+Thirty-fifth+adult+lung+and+heart-lung+transplant+report-2018;+focus+theme:+Multiorgan+transplantation.">The international thoracic organ transplant registry of the international society for heart and lung transplantation: Thirty-fifth adult lung and heart-lung transplant report-2018; focus theme: Multiorgan transplantation.</a> The Journal of Heart and Lung Transplantation: The Official Publication of the International Society for Heart Transplantation. 37 (10), 1169-1183 (2018).</li><li>Valapour, 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=OPTN/SRTR+2017+annual+data+report:+Lung.">OPTN/SRTR 2017 annual data report: Lung.</a> American Journal of Transplantation: Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons. 19, 404-484 (2019).</li><li>Klein, A. 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=Organ+donation+and+utilization+in+the+united+states,+1999-2008.">Organ donation and utilization in the united states, 1999-2008.</a> American Journal of Transplantation: Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons. 10 (4), 973-986 (2010).</li><li>Kotecha, 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=Continued+successful+evolution+of+extended+criteria+donor+lungs+for+transplantation.">Continued successful evolution of extended criteria donor lungs for transplantation.</a> The Annals of Thoracic Surgery. 104 (5), 1702-1709 (2017).</li><li>Singh, 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=Sequence+of+refusals+for+donor+quality,+organ+utilization,+and+survival+after+lung+transplantation.">Sequence of refusals for donor quality, organ utilization, and survival after lung transplantation.</a> The Journal of Heart and Lung Transplantation. 38 (1), 35-42 (2019).</li><li>Cypel, 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=Normothermic+ex+vivo+lung+perfusion+in+clinical+lung+transplantation.">Normothermic ex vivo lung perfusion in clinical lung transplantation.</a> The New England Journal of Medicine. 364 (15), 1431-1440 (2011).</li><li>Wallinder, 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=Early+results+in+transplantation+of+initially+rejected+donor+lungs+after+ex+vivo+lung+perfusion:+A+case-control+study.">Early results in transplantation of initially rejected donor lungs after ex vivo lung perfusion: A case-control study.</a> European Journal of Cardio-Thoracic Surgery: Official Journal of the European Association for Cardio-Thoracic Surgery. 45 (1), 40-44 (2014).</li><li>Sage, 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=Lung+transplantation+from+initially+rejected+donors+after+ex+vivo+lung+reconditioning:+The+french+experience.">Lung transplantation from initially rejected donors after ex vivo lung reconditioning: The french experience.</a> European Journal of Cardio-Thoracic Surgery: Official Journal of the European Association for Cardio-Thoracic Surgery. 46 (5), 794-799 (2014).</li><li>Valenza, 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=Extracorporeal+lung+perfusion+and+ventilation+to+improve+donor+lung+function+and+increase+the+number+of+organs+available+for+transplantation.">Extracorporeal lung perfusion and ventilation to improve donor lung function and increase the number of organs available for transplantation.</a> Transplantation Proceedings. 44 (7), 1826-1829 (2012).</li><li>Fildes, J. 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=Clinical+outcome+of+patients+transplanted+with+marginal+donor+lungs+via+ex+vivo+lung+perfusion+compared+to+standard+lung+transplantation.">Clinical outcome of patients transplanted with marginal donor lungs via ex vivo lung perfusion compared to standard lung transplantation.</a> Transplantation. 99 (5), 1078-1083 (2015).</li><li>Cypel, 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=Experience+with+the+first+50+ex+vivo+lung+perfusions+in+clinical+transplantation.">Experience with the first 50 ex vivo lung perfusions in clinical transplantation.</a> The Journal of Thoracic and Cardiovascular Surgery. 144 (5), 1200-1206 (2012).</li><li>Clark, S. 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=A+new+porcine+model+of+reperfusion+injury+after+lung+transplantation.">A new porcine model of reperfusion injury after lung transplantation.</a> Laboratory Animals. 33, 135-142 (1999).</li><li>Karimi, 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=Technical+pearls+for+swine+lung+transplantation.">Technical pearls for swine lung transplantation.</a> Journal of Surgical Research. 171, 107-111 (2011).</li><li>Kruger, 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=Porcine+pulmonary+auto-transplantation+for+ex+vivo+therapy+as+a+model+for+new+treatment+strategies.">Porcine pulmonary auto-transplantation for ex vivo therapy as a model for new treatment strategies.</a> Interactive CardioVascular and Thoracic Surgery. 23, 358-366 (2016).</li><li>Mariscal, 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=Pig+lung+transplant+survival+model.">Pig lung transplant survival model.</a> Nature Protocols. 13, 1814-1828 (2018).</li><li>Aboelnazar, N. 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=Negative+pressure+ventilation+decreases+inflammation+and+lung+edema+during+normothermic+ex+vivo+lung+perfusion.">Negative pressure ventilation decreases inflammation and lung edema during normothermic ex vivo lung perfusion.</a> The Journal of Heart and Lung Transplantation: The Official Publication of the International Society for Heart Transplantation. 37 (4), 520-530 (2018).</li></ol>

该协议涉及几个关键的手术步骤，需要进行故障排除以确保移植和肺部评估成功。与成年人的肺相比，幼年猪肺非常脆弱，因此手术外科医生在处理猪肺时必须小心。在进行 12 小时的 ESLP 后尤其如此，因为器官将承担液体量并且容易因过度操作而受伤。任何过大的压力都会导致实验肺不张或创伤，从而影响评估结果。同样，幼猪的血管结构非常脆弱。避免 PA 夹扭转至关重要，因为这会导致组织?...

肺移植是终末期肺病的首要长期治疗方法。全世界每年进行超过 4,600 例肺移植手术 1.然而，肺移植目前有很大的局限性。首先，器官的必要性继续使可用的捐赠者黯然失色。尽管自 2012 年以来，由于更多移植候选者被列入名单、供体数量增加以及恢复器官使用得到改善的综合影响，肺移植率每年都在上升，但移植候补名单的死亡率并未显着下降2.器官质量问题是另一个主要限制，据报道器官利用率低至 20%-30%3,4,5。最后，肺移植术后结局的趋势不尽如人意，长期移植和患者结局仍落后于其他实体器官移植2.
一种新兴技术，即非原位肺灌注（ESLP），有可能减轻这些局限性。ESLP 已越来越多地被用作评估和修复边缘供体肺的方法，并且已证明移植扩展标准供体 （ECD） 肺6、7、8、9、10 后可接受的短期和长期结果。因此，ESLP 将一些中心的利用率提高了 15%-20%6、7、8、9、10、11。
适当的ESLP研究需要对体外研究结果进行体内验证;然而，关于 ESLP12、13、14、15 的猪肺移植模型的文献有限。此外，现有文献提供的关于约克郡猪肺移植麻醉管理的细节不足，肺移植的血流动力学可能非常不稳定12,13,14,15。建立一个易于重复的模型将允许对当前的ESLP策略进行体内验证，并对各种干预措施进行临床前评估，以减少肺缺血再灌注损伤。本研究的目的是描述用于 ESLP 的原位左肺同种异体移植的猪模型。该方案包括麻醉剂和手术技术的描述、自定义手术清单以及有关故障排除体验和方案修改的详细信息。本文还讨论了左肺猪移植模型的局限性和益处。本手稿没有概述35-50公斤约克郡猪猪肺的检索过程，也没有涵盖ESLP的建立和终止。该协议专门针对受体移植手术。

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			<td>21-Gauge Needle</td>
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left lung orthotopic transplantation in a juvenile porcine model for eslp

肺移植是终末期肺病的金标准治疗方法，全世界每年进行超过 4,600 例肺移植手术。然而，肺移植受到可用供体器官短缺的限制。因此，候补名单的死亡率很高。在一些中心， 非原位 肺灌注 （ESLP） 使供体肺利用率提高了 15%-20%。ESLP 已被应用为评估和修复边缘供体肺的一种方法，并已证明移植扩展标准供体 （ECD） 肺后可接受的短期和长期结果。需要大型动物（体内）移植模型来验证正在进行的 体外 研究结果。人类和猪之间的解剖学和生理学差异带来了重大的技术和麻醉挑战。易于复制的移植模型将允许对当前的ESLP策略进行体内验证，并对旨在改善供体肺功能的各种干预措施进行临床前评估。该方案描述了原位左肺同种异体移植的猪模型。这包括麻醉和手术技术、定制的手术清单、故障排除、修改以及该方法的优点和局限性。

所有结果均在 NPV-ESLP16 12 小时后再灌注 4 小时的背景下进行。在肺外植体期间，有几种临床结果需要预期（图3）。通常，猪在左肺移植成功后将保持血流动力学稳定，但由于对手术的血管舒张反应，可能需要低剂量输注去氧肾上腺素（剂量范围：2-10 mg/h）。心率目标应约为 100-120 次/分，SpO 2 > 90% 时呼吸频率 （RR） 为 8-30%，平均动脉压 （MAP） > 60 m...

Watch this Scientific Journal Video about Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP at JoVE.com

Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP

该协议描述了设计用于ESLP研究的原位左肺同种异体移植的幼猪模型。重点是麻醉和手术技术，以及关键步骤和故障排除。

ESLP幼猪模型中的左肺原位移植

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. ...

该方案是大型动物模型最直接的肺移植方案之一，在手术管理方面提供了最详细的信息。该技术的主要优点是描述手术，评估移植后孤立的左肺功能，并在手术期间稳定猪。该方法有利于非原位肺灌注研究，以验证体内再灌注移植模型的异位发现。肺移植的体内再灌注模型有利于测试非原位肺充注结果，也可用于人工器官测试，例如生物工程或机械肺。首先将猪置于右侧褥疮位置，进行左前外侧开胸手术。然后用触诊识别剑突左肩胛骨的尖端，低于胸骨，并将两个识别的斑点连接起来，标记20厘米长的开胸切口部位。麻醉后，用手术刀在皮肤上做一个切口，然后用电烙术解剖动物的皮下层和肌肉层。背阔肌必须分开。在确定切口正下方的肋骨后，在肋骨顶部烧灼以暴露肋间肌肉，同时避开肋间神经血管束。使用蚊子止血器刺穿肋骨正上方的肋间肌肉，然后用手指感觉胸部内部是否有粘连。在烧灼肋骨上缘的同时，使用 Yankauer 抽吸将肺推开，将开胸手术向前延伸至距胸骨一英寸，并向后延伸至椎旁肌肉。通过插入 Cooley 胸骨牵开器，打开 10 厘米宽的开胸手术以缩回肺部并暴露左合子静脉。使用 Metzenbaum 剪刀和细下部沿圆周解剖左半合子静脉。通过用丝带绕血管，结扎并横断静脉。解剖左肺动脉（PA）和左肺静脉（PV），然后用丝带环绕静脉。肝素给药五分钟后，用DeBakey十字夹夹夹住PA，用Satinsky夹夹夹左前PV，用勺罐夹夹左支气管。横断 PA、左下 PV 和左支气管，留下至少 0.5 厘米的组织袖带。分离左下肺韧带后，切除左肺。移植时，将供体肺插入受体的胸部，从下叶开始，不要强迫肺进入该位置。然后使用 TF 针上的 4.0 Prolene 缝合线进行支气管吻合术。为了评估抽吸分泌物中的支气管吻合，使用适配器连接将支气管镜插入气管插管进行左肺和右肺支气管镜检查，然后在将支气管镜推进左支气管镜进入左支气管之前将内窥镜连接到抽吸。观察肺呼吸，检查支气管吻合术的生命体征。在右侧重复该过程。接下来，在 BV-1 针上使用 6.0 Prolene 缝合线进行端到端左心房或 LA 吻合术，然后修剪多余的组织。将供体 SVP 并入下 PV 和 LA 吻合术后，使用运行的端到端吻合术在 BV-1 针头上用 6.0 Prolene 缝合线完成 PA 吻合术。之后，确认肝素化并给动物施用钾转移。然后部分打开 PA 夹进行脱气，让血液流过缝合线。接下来，松开 LA 夹，让缝合线回流，以进一步释放任何滞留的空气。系好 LA，然后取下 PA clamp 完全。完成后，取下支气管钳夹并增加潮气量，以达到每公斤 10 毫升的目标。插入 20 法式可塑胸管后，分三层关闭开胸手术。使用 Metzenbaum 剪刀打开左胸膜，从左下叶采集血样。打开右胸膜，解剖右肺动脉。然后放置夹子以停止右肺灌注。将一根 21 号针引向左肺静脉并远离左心房，从 LA 吻合口采集血液样本以评估左肺功能。夹紧后，用 21 号针头朝向左肺，从左侧 PV 吻合口处采集血样。代表性分析表明，在肺移植方案中，典型的PF比值变化和水肿形成。在再灌注时，PF比值下降了约100毫米汞柱，因为左肺在氧合时不能立即有效。在4小时的离体左肺评估中，PF比值稳定。在成功的移植中，由于循环中残留的血液，左肺的体重可能会增加约 20%至 50%。左肺移植12 h后，进行血气分析。观察到在60至120分钟的再灌注期间钾水平增加。在四小时的再灌注期间，大约需要两到四个班次，以将钾保持在每升 5 毫摩尔以下。侧咬夹可用于血管控制，并且可以很容易地缝合血管，而不是缝合受体的上肺静脉。

Research

JoVE Journal

Medicine

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

闭塞性细支气管炎在原位肺移植小鼠模型的发展

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

科研

医学

Orthotopic Small Bowel Transplantation in Rats

大鼠原位小肠移植

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Liver Transplantation in Rats

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

部分原位肝移植大鼠模型的肝动脉重建的外科手术

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

心肌缺血再灌注损伤的通过左前降支结扎小鼠模型

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

使用Active波尔图腔静脉分流术猪肝脏采购和原位移植技术

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

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 ...

Orthotopic Hind Limb Transplantation in the Mouse

原位后肢移植中的鼠标

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

正交心脏移植的临床前猪模型

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...