Name: a pre-clinical porcine model of orthotopic heart transplantation
Uploaded: 2019-04-27T14:30:00
Description: Watch this Scientific Journal Video about A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation at JoVE.com

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机构动物护理委员会批准了所有实验规程,动物按照国家研究理事会实验室动物资源研究所编写的《保护和使用实验室动物指南》进行了处理。雄性约克郡猪(40-50公斤)用于进行正交心脏移植(动物大小可能因调查人员的判断力和实验目标而异)。
1. 捐助者程序
<ol><li>麻醉诱导和动物制备:<ol>
<li>使用氯胺酮(20毫克/千克)、米达佐兰(0.3毫克/千克)和阿托品(0.04毫克/千克)的肌肉注射?...

<ol>
	<li>Lund, L. H., Edwards, L. 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=The+Registry+of+the+International+Society+for+Heart+and+Lung+Transplantation:+Thirty-third+Adult+Heart+Transplantation+Report-2016;+Focus+Theme:+Primary+Diagnostic+Indications+for+Transplant.">The Registry of the International Society for Heart and Lung Transplantation: Thirty-third Adult Heart Transplantation Report-2016; Focus Theme: Primary Diagnostic Indications for Transplant.</a> The Journal of Heart and Lung Transplantation. 35 (10), 1158-1169 (2016).</li><li>Lund, L. H., Edwards, L. 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=The+Registry+of+the+International+Society+for+Heart+and+Lung+Transplantation_+Thirty-first+Official+Adult+Heart+Transplant+Report-2014;+Focus+Theme_+Retransplantation.">The Registry of the International Society for Heart and Lung Transplantation_ Thirty-first Official Adult Heart Transplant Report-2014; Focus Theme_ Retransplantation.</a> The. Journal of Heart and Lung Transplantation. 33 (10), 996-1008 (2014).</li><li>Cos&#237;o Carmena, M. D. G., G&#243;mez Bueno, 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=Primary+graft+failure+after+heart+transplantation:+characteristics+in+a+contemporary+cohort+and+performance+of+the+RADIAL+risk+score.">Primary graft failure after heart transplantation: characteristics in a contemporary cohort and performance of the RADIAL risk score.</a> The Journal of Heart and Lung Transplantation. 32 (12), 1187-1195 (2013).</li><li>Fedak, P. W. 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本手稿描述了一个大型动物正交心脏移植的临床前模型。各种异位心脏移植的小型动物模型已成功地用于研究新疗法对改善器官保存和减少缺血再灌注损伤的效果11、12、 13.此外,小型动物模型允许生存模型和长期随访,以调查排斥和心脏异体血管病的发展11,12,13。<sup class="xref"...

在第一次成功报告50年后,心脏移植仍然是符合资格的晚期心力衰竭患者的黄金标准治疗。虽然长达4小时的缺血时间是耐受的,但超过6小时的缺血时间与劣质结果2有关。原发性移植功能障碍仍然是移植2、3后早期发病和死亡的主要原因。原发性移植功能障碍的病因是多因素的,包括使用边缘器官、受体肺血管疾病、高急性排斥和在移植时遭受的缺血再灌注损伤3。
多项研究已研究新的方法,供体心脏保存,以减少原发性移植功能障碍的发生率4,5,6,7。评估缺血性灌注损伤或异位心脏移植的小鼠模型中的新技术和治疗方法是常见的做法。此外,小动物模型允许生存模型和长期随访,以调查排斥和心脏异体血管病11,12,13的发展。然而,这些策略大多未能通过初步的临床试验,或从未达到这一阶段。在临床相关的大动物模型中评估新疗法,以便有效可靠地翻译基础研究的发现,这一点至关重要。
使用大型动物模型时,猪心通常被认为是与人类心脏在解剖学上最相似的。因此,它是进行心脏外科研究的理想平台。然而,在使用猪模型时,需要考虑几个重要因素。首先,组织通常被描述为脆弱和易碎,特别是在右中庭和肺动脉,容易撕裂14。此外,猪心脏被认为对操纵敏感,容易出现心律失常,这就是为什么在实验开始时应该定期对每只动物进行抗动脉炎。猪模型和临床心脏移植之间的一个重要解剖区别是猪的左血脉,直接进入冠状动脉。这必须在接受者过程中连接,以避免连续出血。最后,猪模型对缺血非常敏感,但仍适用于心脏移植的急性研究。
本手稿描述了一个临床前大动物(猪)的正交心脏移植模型,该模型已经牢固建立,并用于研究新的心脏保护策略5,6,8 ,9.

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a pre-clinical porcine model of orthotopic heart transplantation

在第一次成功报告发布五十年后,心脏移植仍然是符合资格的晚期心力衰竭患者的黄金标准治疗。多种小动物心脏移植模型已用于研究新疗法的急性和长期影响。然而,很少在临床试验中进行测试和证明成功。在临床相关的大动物模型中评估新疗法,以便有效可靠地翻译基础研究的发现,这一点至关重要。在这里,我们描述了一个临床前大动物(猪)的正交心脏移植模型,该模型已经牢固建立,并以前用于研究新的心脏保护策略。该程序侧重于急性缺血再灌注损伤,是研究在较小的实验模型(如鼠害模型)中经过测试和验证的新型干预措施的可靠方法。我们证明了它在评估移植后早期的心脏性能以及该模型启用的其他潜在可能性方面的有用性。

这种临床前模型自1994年以来已成功使用5,6,8,9。表1展示了在基线上采取的压力-体积关系和超声心动图参数的代表性结果,以及一组5个实验中3小时移植后的结果。虽然我们看到移植后肌体收缩性下降,但在统计学上并不显著。
图 1显示了从同一个?...

Watch this Scientific Journal Video about A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation at JoVE.com

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

在这里,我们描述了一个临床前大动物(猪)的正交心脏移植模型,该模型已经牢固建立并用于研究新的心脏保护策略。

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 ...

使用临床相关的大型动物模型评估新的心脏保存策略对于可靠的临床翻译非常重要。猪心模型用于心脏外科的研究是一个非常有用的模型，因为人类和猪的心脏在解剖学、生理学、甚至大小方面有很多相似之处，因此它是一个几乎理想的研究平台。打开麻醉供体猪的腹孔后，请助手小心地将右心室流出道和左下角的肺动脉缩回，并小心地将主动脉缩回右侧，以避免肺动脉直接病变。使用烧灼解剖大动脉肺空间，从结缔组织中清除提升大动脉的前部。小心地缩回右心室流出道后，使用 4.0 PROLENE 缝合线将钱包串缝放在近近上升大母蛋白出现处，并使用图面线固定缝合线。将心痛分娩管插入钱包串缝合内的上升大母，并拧紧图条，以确保管。接下来打开左劣肺静脉和上优和劣等的静脉，以确保足够的心脏通气，并放置大动脉交叉夹在开界上升大动脉上。启动心痛输液，针对 80 至 100 毫米汞的大音根压力。将0.9%氯化钠冰泥涂抹在胸腔和器官上。当心痛输液完成后，以传统方式进行切除术，在主动脉和肺动脉后以及分叉后切开主动脉，以确保植入的足够长度。要在接受者中启动心肺旁路，首先使用 Metzenbaum 和直角钳子解剖上优的静脉冠状动脉和下一层静脉动脉与劣质的静脉道和麻醉接受动物的胸腔。单独用 0 丝领带包围上等的维纳卡瓦，用旅游布固定每盘胶带。使用 4-0 PROLENE 缝合线将两个同心钱包串缝合线放在远角大肠的出入上，并在心向反射的水平上将钱包串缝合线放在下级和优越的 vena cavae 上。用图片固定每个缝合线，并使用塞尔丁格技术将17-21法国动脉管插入大动脉。使用 3/8-3/8 连接器将管与旁通电路的动脉线连接。在放在维纳卡瓦纳的钱包串缝合线的中心做一个五毫米的切口，用一个小角度的仪器将切口扩张。使用24-28法式右角，单阶段静脉管，可以拉下优势和劣等的维纳卡瓦，在优越的维纳卡瓦和劣质的维纳卡瓦，远离心脏的角度。拧紧手持钱包串缝合线的图尼奎特，以固定管，并使用 3/8-3/8 1/2-Y 连接器将管弦连接到旁通电路的静脉线。然后启动心肺旁路。心肺旁路启动后，使用锋利的解剖仪器和直角包围左血红酶静脉，用 O-silk 领带将血管绕开。交叉夹紧接受者大动脉紧直到动脉管和圈套两个维纳卡瓦与以前放置的O-silk领带。比较捐赠者和接受者的左心房袖口尺寸，并根据需要修改每个大小。使用先前放置的心肺痛管将第一个心痛剂量送到供体心脏。接下来使用标准的比亚审原蛋白技术和SH针，在左中庭和下级维纳卡瓦的右下缘交界处放置一个4-0PROLENE缝合线。将另一个缝合线从第一个缝合线180度处放置，连接供体和接受者袖口，然后完成从上一缝合线到下级缝合的后壁前壁。对于右中庭，将捐赠者右中庭从附属物打开至劣质的 Vena 卡瓦，创建与接受者袖口大小相匹配的捐赠者袖口，然后从劣质的 Vena 卡瓦和右中庭之间的交界处开始，完成内壁肛门病，然后完成侧壁。对于肺动脉，修剪接受者和供体肺动脉的边缘以创建匹配的大小。使用 BB 针将 5-0 PROLENE 缝合线放在左侧壁上，将供体和接受者血管联合，在右侧边缘放置另一根 PROLENE 缝合线。然后完成劣质壁性腺体病和前壁性结膜病。对于大母体，修剪边缘，就像刚刚演示的一样，并在连接供体和接受者血管的左面壁上放置缝合线。然后完成劣质壁和前壁脑膜炎，然后启动心肺旁路60分钟的再灌注。为了对移植物进行功能评估，在劣质的 Vena cava 周围放置一个 0 丝领带，并通过一个小的腹腔心室插入压力体积导管，以便连续测量左心室压力-体积关系。这里具有代表性的结果来自压力-体积关系和超声心动图参数在基线，并在移植后三小时在一组五个实验显示。在稳态评估期间，会记录与体积相关的参数，例如发达压力的最大速率和最小速率。通过间歇性遮挡低射的 Vena cava 获得与体积无关的参数，在此期间，左心室的体积逐渐减小，并记录最终收缩和结束舒张压力体积关系。预加载可招募冲程工作数据可用于确定冲程工作与相应的最终舒张体积之间的关系。各种其他代谢参数也可以用这个模型测量。此过程需要至少一名训练有素的外科医生和大约五个实践实验，以优化每个研究组的研究方案。猪的心脏脆弱，易碎，特别敏感操纵和缺血，因此容易流泪。重要的是要修改所有的麻醉，以确保最小或无出血。该协议可以帮助开发新的心肌保存策略，以减少缺血灌注损伤，并防止原发性移植失败。

Research

JoVE Journal

Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

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 ...

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

酪氨酸激酶抑制剂的临床前评价急性白血病的治疗

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

用改进的袖套法小鼠颈部心脏移植模型

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

肺动脉瓣使用异位心脏移植小鼠模型移植

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

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 ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

人前列腺癌的临床前原位小鼠模型

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Pre-clinical Model of Cardiac Donation after Circulatory Death

循环死亡后心脏捐献的临床前模型

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

重建显微外科缺血再灌注损伤研究的临床前大鼠模型

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

使用24小时器官保存和连续遥测的猪模型中正交肾自动移植

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...