CT检查支持由瑞士国家科学基金会（分数批3232230-126233）。这项研究是支持的Artères基金会，阿斯特拉基金会欧洲和Boninchi的基金会。

1。捐助行动<ol><li>麻醉雄性大鼠，体重200±20克异氟醚与锥上的面具（归纳为3％，2％，在操作过程中，1升/分钟空气流量，FIO2 70％）。 </li><li>执行一个大型的中位数和横向剖腹探查术和灸两侧上腹船只。 </li><li>切断镰状韧带，左膈静脉分开的suprehepatic腔静脉。最后，分成两个7-0丝结扎膈肌静脉。 </li><li>切断左侧的三角形和胃肝韧带。 </li><li> 7-0结扎（凝血是一种有效的替代品）之间的分化肝食管韧带和动脉。 </li><li>隔离左肾静脉下腔静脉infrahepatic。分离右肾静脉和周围组织之间划分10-0和7-0结扎，其近端和远端的两端，分别。 </li><li>除以右肾上腺静脉之间7-0 ligatur，ES和自由的肝切割温柔的牵引下，从后韧带。 </li><li>隔离gastrosplenic静脉和分裂之间的10-0和7-0结扎，近端和远端的两端，分别。 </li><li> 7-0关系之间的划分适当的肝动脉。 </li><li>分离出的胰十二指肠静脉和10-0和7-0结扎，其近端和远端，分别划分。 </li><li>通过实践在1厘米的距离从肝门的一个小切口插入胆总管1 22G，3.5毫米的支架。固定支架在7-0结扎的位置。 </li><li> ，通过阴茎背静脉注入10的用户界面，为1毫升生理盐水稀释肝素。 </li><li> Cannulate腔PORTA（尽可能从肺门），与21G针轻轻冲洗肝脏冷乳酸林格氏液（或其他保存液）20毫升。在同一时间，减少低于肾静脉腔静脉，使足够outfl唔。肝脏冷灌注应持续一两分钟之间。 </li><li>切断胰十二指肠静脉下腔PORTA。 </li><li>切断胆总管远端结扎支架周围的。 </li><li>完成的infrahepatic腔静脉切开。 </li><li>切肝上腔静脉脱脂隔膜取出肝脏。 </li><li>放入充满了寒冷的乳酸林格氏液，躺在一个冰垫盆地盆地的肝脏。 </li></ol> 2。移植物的制备<ol><li>其袖口插入腔PORTA，然后船只周围边缘外翻，暴露内膜。争取在7-0结扎袖口外翻段。 </li><li>上腔静脉infrahepatic重复同样的操作。 </li><li>对下腔静脉infrahepatic近端部分微钳（4-6毫米长）的位置。此操作可避免血液门户再灌注后的损失。 </li><li>推翻肝脏和插入两个8-0的普理灵缝线肝上下腔静脉，从外到内的另一侧边缘。 </li><li>存放在冰箱中的移植（成乳酸林格氏液浸泡），在0-4°C。 </li></ol> 3。受体手术<ol><li>麻醉雄性大鼠，体重200±20克异氟醚与锥上的面具（归纳为3％，2％，在操作过程中，1升/分钟空气流量，FIO2 70％），并保持在一个温暖的垫。值得注意的是，应符合受援国和捐助国的重量（​​±40克）。 </li><li>注入10毫升的生理盐水皮下注射，哌拉西林/他唑巴坦肌肉注射0.03Ğ前剖腹手术。 </li><li>执行中线施福建耻骨剖腹。 </li><li>自动静电镊子扭转的剑突过程中为了更好地暴露肝脏。 </li><li>削减前韧带和两个7-0关系划分左膈静脉。 </li><li> 7-0结扎（混凝之间的分化肝食管韧带和动脉n是一个有效的替代）。不显示视频（捐助者的操作相同）。 </li><li>隔离的infrahepatic腔静脉，右肾静脉。 </li><li> 7-0连字和自由切割温柔的牵引下，从肝后韧带（在视频显示（捐助者的操作相同））之间的分割右肾上腺静脉。 </li><li>放置周围胆总管7-0结扎，仅低于其分裂。把另一个结扎1毫米削减之间。 </li><li>隔离肝动脉和脐划分7-0连写。 </li><li>轻轻分开左，右门静脉分支。注意不要损害肝动脉或胃脾静脉。 </li><li>定位颅右肾静脉腔腔静脉周围环，。然后保护血管壁，环，把坚持四项基本原则9-0缝线。 </li><li>门户环上腔PORTA重复同样的操作。 </li&gt; <li> ，通过阴茎背静脉注入10的用户界面，为1毫升生理盐水稀释肝素。 </li><li>设置麻醉和供氧浓度100％，0.25％。 </li><li>钳略高于右肾静脉下腔静脉infrahepatic，然后钳上面的胃脾静脉腔PORTA。最后夹住肝上腔静脉，一定要包括至少2毫米膜片。 </li><li>切肝上下腔静脉紧密的实质。 </li><li>削减的左边和右边的门静脉分支和分裂之间的隔膜。 </li><li>削减的infrahepatic腔静脉密切的实质，不切开任何先前定位缝线关注。取出肝脏放入腹腔移植。 </li><li>执行的肝上下腔静脉移植和受援国之间的端至端吻合。启动与运行由左到右后壁缝合，然后进行前壁由右至乐英尺保持直到结束前缝合线松动，为了让前拧紧两端的内部腔慷慨潮红。 </li><li>插入到门户的环柄尾分行的逼近，然后将其插入腔移植PORTA寻找到插槽的位置，与门户袖口，缝颅分支的逼近。 </li><li>关闭的逼近，同时用生理盐水冲洗收件人的腔PORTA和袖口允许插入的船只。争取用7-0周的领带腔围绕其袖口收件人的PORTA。 </li><li>删除的逼近。为了打开钳对肝上下腔静脉和腔PORTA:门户网站流量正在重建。 </li><li>取出9-0连字切割额外罚款剪刀的门户环。 </li><li>插入腔环柄尾分行的逼近，然后插入颅缝腔静脉移植infrahepatic<html>分公司的逼近，与下腔静脉入插槽找到它的位置的袖口。 </li><li>关闭的逼近，同时用生理盐水冲洗收件人的infrahepatic腔静脉和袖口允许插入的船只。争取用7-0周的领带收件人围绕其袖口腔静脉。 </li><li>删除的逼近。为了打开下腔静脉移植和收件人的infrahepatic microclamps:静脉血流正在重建。 </li><li>执行部分收件人的胆总管切口，适用于温柔的牵引，向头和插入以前插入到移植的胆管支架自由端。圆周7-0领带固定支架。 </li><li>关闭层用5-0缝线的动物。允许从觉醒的自由水和食物。 </li><li>根据当地机构协议管理的有效镇痛。一个24小时的最低期限的建议。 </li></ol>_content"&gt;注:年底到年底可以进行肝动脉吻合，当需要通过实验设计（动脉大鼠肝移植后长期生存的需要）。 4。代表结果所描述的技术可以成功地掌握我们的团队，经过约10个训练手术。它允许长期100％（&gt; 21天）连续36刘易斯，刘易斯和黑刺鼠到刘易斯移植的生存，经过短期培训期间（同基因肝移植受者的数量一直维持存活超过12个月）。同基因移植后第1天肝功能试验表明:天门冬氨酸转氨酶（AST）的中位数:84 U / L（51.7 - 92.7）和中位数丙氨酸转氨酶（ALT）:172.5 U / L（127.5 - 240.2）。平均无肝期（从腔PORTA夹紧移植再灌注）为14±2分钟，最长的肝上下腔静脉吻合的步骤，（运行小号uture）时间9±2分钟。 快速连接器系统允许1.55毫米口径袖口腔PORTA和2.40毫米口径袖口上腔静脉（体重200±20克的收件人）的定位。相反，当我们试图移植用镰田的技术，他们可以不超过1.40毫米，分别2.16毫米（应变和体重匹配的大鼠）。快速链接环， 如图7所示，旨在保持最佳伸展收件人的船只，允许最低卡尺和长度的浪费，导致在接近生理的血 ​​流动力学结果。 <img src="/files/ftp_upload/4143/4143fig1.jpg" alt="图1" /> 图1。戒指措施。 <img src="/files/ftp_upload/4143/4143fig2.jpg" alt="图2" /> 图2。Microclamps措施。 <img src="/files/ftp_upload/4143/4143fig3.jpg" alt="图3" /> 图3。快速交联剂套件。 <img src="/files/ftp_upload/4143/4143fig4.jpg" alt="图4" /> 图4。快速链接武装的。 <img src="/files/ftp_upload/4143/4143fig5.jpg" alt="图5" /> 图5。快速链接关闭。 <img src="/files/ftp_upload/4143/4143fig6.jpg" alt="图6" /> 图6。SHVC钳。 <img src="/files/ftp_upload/4143/4143fig7.jpg" alt="图7" /> 图7。经典与快速连接器的辅助袖口吻合血流动力学。 <img src="/files/ftp_upload/4143/4143fig8.jpg" alt="图8" /> 图8。捐赠者和接受者的皮肤准备切口。

<ol>
	<li>Starzl, T. E., Marchioro, T. L., Porter, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+and+clinical+observation+after+homotransplantation+of+the+whole+liver.">Experimental and clinical observation after homotransplantation of the whole liver.</a> Revue internationale d'hepatologie. 15, 1447 (1965).</li><li>Starzl, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+liver+transplantation.">Evolution of liver transplantation.</a> Hepatology. 2, 614S-636S (1982).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47 (1979).</li><li>Kashfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplantation proceedings. 37, 185-188 (2005).</li><li>Hori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+and+innovative+techniques+for+liver+transplantation+in+rats:+A+surgical+guide.">Comprehensive and innovative techniques for liver transplantation in rats: A surgical guide.</a> World Journal of Gastroenterology: WJG. 16, 3120 (2010).</li><li>Oldani, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+technique+for+rat+liver+transplantation+using+Quick+Linker+system:+a+preliminary+result.">A novel technique for rat liver transplantation using Quick Linker system: a preliminary result.</a> Journal of Surgical Research. 149, 303-309 (2008).</li><li>Nosaka, T., Bowers, J. L., Cay, O., Clouse, M. E. <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+in+rats.">Biliary complications after orthotopic liver transplantation in rats.</a> Surg. Today. 29, 963-965 (1999).</li><li>Li, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+incidence+of+biliary+complications+in+rat+liver+transplantation:+can+we+avoid+it.">High incidence of biliary complications in rat liver transplantation: can we avoid it.</a> World J. Gastroenterol. 17, 3140-3144 (2011).</li><li>Peng, Y., Gong, J. P., Yan, L. N., Li, S. B., Li, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+two-cuff+technique+for+orthotopic+liver+transplantation+in+rat.">Improved two-cuff technique for orthotopic liver transplantation in rat.</a> Hepatobiliary Pancreat. Dis. Int. 3, 33-37 (2004).</li><li>Urata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+survival+in+rat+liver+transplantation+with+extended+cold+preservation:+role+of+portal+vein+clamping+time.">Decreased survival in rat liver transplantation with extended cold preservation: role of portal vein clamping time.</a> Hepatology. 28, 366-373 (1998).</li></ol>

没有利益冲突的声明。

目前肝移植模型，可以方便地建立。几个关键点在我们的经验，有助于减少死亡的危险（≤20％）。早期死亡，再灌注前三个小时内发生的，是最常见的原因到肠梗塞，作为长期夹紧门户的后果。因此，建议保持在无肝期15分钟的理想限值（20分钟还是可以接受的）。中央空调等症也可以是突然死亡的原因，它是强制刷新之前的所有静脉吻合。 术后第二天一和三之间发生的死亡往往是由于肝功能衰竭或下腔静脉血栓形成。因此，应该避免任何侵略性移植前捐助explantation（我们用20毫升冲洗约60秒）冲洗，并确保适当的低温保存（为0-4°C间乳酸林格氏液充分浸泡）。血栓形成的风险可降低减少内膜的操作和维护其完整性。我们不建议手术后使用的抗凝。 术后第四天九之间的死亡通常与细菌性胆管炎7。根据我们的经验，piperacilline /他唑巴坦（0.03克）的单剂量给予前剖腹是足够的，以尽量减少胆总管感染的风险。 死亡后10天是罕见的事件。然而，由于动脉血流的情况下，一些胆管的问题，可以观察到8 cholastasis。根据研究者的需要，动脉可能被视为9。值得注意的是，冷缺血时间长达24小时，可以被视为可以接受的10。 快速连接器系统的最初目的是进行肝移植三袖套吻合（很短的无肝期）。然而，三袖套TEchnique需要使用非常小的卡钳为肝上下腔静脉（SHVC）袖口，它并没有被广泛接受，由于11-14肝脏流出问题的风险。因此，我们主张快速链接辅助三袖套法只有当需要热缺血时间短十多分钟。但在大多数研究中，植入阶段15-18分钟是可以接受的，允许使用两袖套法，更生理SHVC排水。在这个视频文件所示，我们经常使用快速连接器系统，在第二部分仅植入。虽然一些额外的手术时间需要快速连接器手柄的位置，以最小的风险有关这个过程的一部分，并可以缩短无肝期的关键得益于容易袖口插入。快速连接器系统的其他优点是一个更好的移植受体血管对齐，并使用更多更好的血流动力学结果袖口相比先前所描述的技术（包括镰等人描述的。）6（ 图7）。

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orthotopic liver transplantation in rats

在肝移植领域的临床进展已基本支持1,2动物模型。镰等。在1979年出版以来的第一次原位肝移植，这种模式仍然是金标准，尽管各拟议的替代技术。然而，它的广泛使用是有限的，其陡峭的学习曲线5。 在这个视频文件，我们显示了镰田的两袖套法简单，易于建立的修订。肝上下腔静脉吻合手动执行与运行缝合，静脉PORTA和infrahepatic的下腔静脉吻合，利用快速链接袖口系统6。制造业的快速连接器套件显示在一个单独的视频文件。

Watch this Scientific Journal Video about Orthotopic Liver Transplantation in Rats at JoVE.com

Orthotopic Liver Transplantation in Rats

我们提出了一个经典的大鼠原位肝移植二袖套技术易于建立的修订。

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 ...

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JoVE Journal

Medicine

29.8K 次观看.  University of Geneva Hospitals. 我们提出了一个经典的大鼠原位肝移植二袖套技术易于建立的修订。

视频: Orthotopic Liver Transplantation in Rats

Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

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 ...

科研

医学

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

原位主动脉瓣移植在小鼠血管疾病的研究

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

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

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

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 ...

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

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular 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 ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous perfusion of organs provides the opportunity to improve organ quality and allows ex situ viability assessment of donor livers prior to transplantation. This video article provides a step by step protocol for ex situ normothermic machine perfusion (37 &#176;C) of human donor livers using a device that provides a pressure and temperature controlled pulsatile perfusion of the hepatic artery and continuous perfusion of the portal vein. The perfusion fluid is oxygenated by two hollow fiber membrane oxygenators and the temperature can be regulated between 10 &#176;C and 37 &#176;C. During perfusion, the metabolic activity of the liver as well as the degree of injury can be assessed by biochemical analysis of samples taken from the perfusion fluid. Machine perfusion is a very promising tool to increase the number of livers that are suitable for transplantation.

Ex Situ Normothermic Machine Perfusion of Donor Livers

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

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 ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

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 Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

应用新型磁吻合技术 Suprahepatic 腔静脉重建大鼠原位肝移植模型的研究

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

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 ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

大鼠原位肾移植: 一种新的简化的手术方法

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

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

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

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 ...