这项工作得到了 2022 年 JST 基础研究（日本移植学会）的支持。

该实验方案得到了京都大学动物实验委员会的批准。该研究使用了从商业来源获得的 C57BL/6 小鼠，年龄超过 10 周，体重在 25 g 至 30 g 之间（参见 材料表）。所有动物均用2.5%异氟烷麻醉（遵循机构批准的方案），在特定的无病原体条件下维持，所有实验程序均按照京都大学的《动物实验条例》进行。用于研究的适当仪器列在 材料表 中，如图 1 和 图 2 所示。
1. 动物选择
<ol><li>使用体重为25-30克的小鼠。 注意：虽然小鼠肝脏同种异体移植物2 在主要组织相容性复合物屏障中被普遍接受，但我们在这项研究中选择了同基因 MOLT 模型，以关注冷缺血再灌注损伤10 的详细机制。不建议使用体重小于 25 g 的小鼠，因为不可能将内部支架插入细胆管。同样，由于血管周围存在大量腹内脂肪，因此不建议体重超过 30 克的小鼠。</li>
</ol>2. 袖口制作
<ol><li>分别为 PV 和 IHIVC 准备 20 G 和 16 G 静脉导管。</li>
	<li>使用手术刀切开导管以形成袖带。袖带的主体长度应为2 mm，延伸手柄为1 mm（图3A）。 注意：如果手柄太大，插入袖带可能会变得困难。</li>
	<li>在袖带的表面，使用手术刀刀片的背面创建一个浅凹槽，用于螺纹固定。 注意： 将手术刀刀片的背面贴在袖带上时，请确保刀片的尖端呈大约 60 度角（如图 3B 所示）。制作第一个凹槽后，固定手术刀刀片并旋转袖带。理想情况下，建议使用两个凹槽，但单个凹槽就足够了，没有问题。</li>
</ol>3. 袖带附件
<ol><li>在一个塑料矩形容器中，放置小冰块，并在顶部放置一个小金属杯（直径 6 厘米）。用器官保存液填充杯子（参见 材料表），并将同基因移植物放入其中。</li>
	<li>用棉签轻轻滚动同基因肝移植物（从另一只供体小鼠中提取），确保肝门朝上。用器官保存液彻底清除储存在 IVC 腔中的任何血液。</li>
	<li>使用例如连接到脾静脉的螺纹将 PV 穿过袖带（见 图 4A）。</li>
	<li>将袖带手柄置于 12 点钟位置，用斗牛犬夹固定手柄和袖带amps（参见 材料表），将其放置在距离 PV 边缘 2 至 3 毫米的位置。</li>
	<li>使用大血管镊子和固定模具进一步固定斗牛犬夹以建立手术区域（见 图4B）。</li>
	<li>以大约 15 至 20 倍的放大倍率仔细检查 PV 流明。如有必要，滴水可以提高管腔的可见度。</li>
	<li>轻轻抓住 PV 流腔并通过袖带将其外部化。</li>
	<li>一旦完全外化，使用 8-0 通过双重连接将其固定丝线沿着袖口的凹槽（图4C，D）。 注意： 确保结扎线没有松动。另外，注意结扎点不要太大，因为它可能会干扰袖带插入。结扎点可以定位在任何方向上。</li>
	<li>使用类似于PV的程序将袖带连接到IHIVC（图4E）。 注意：用袖带夹住 IHIVC 时，请避免咬入肝实质。由于颈部比 PV 短，因此请仔细确定夹紧位置。</li>
	<li>通过将线绕在 IHIVC 的底部以暂时连接 IHIVC 来创建一个滑结（图 5）。再灌注后将移除该线。</li>
</ol>4. 门静脉重建
<ol><li>通过用2.5%的异氟烷诱导麻醉来麻醉受体小鼠，并将其降低至0.6%，然后再提取已发表的文献2中所述的天然肝脏。在剖腹手术前，用交替的碘伏和 70% 乙醇对手术区域进行至少 3 次 dinsini 感染。</li>
	<li>确保肝叶位置正确，并且套囊 PV 没有扭结和扭转。</li>
	<li>使用 Pean 镊子轻轻抓住接受者的 PV 边缘，并用钳子将其固定。 注意：在本文中，钳子是指用于将豌豆镊子固定在桌子上的机械装置。有关虎钳的更多信息，请参阅 材料清单。</li>
	<li>将血管钳夹应用于接受者的 PV 并通过 8-0丝线缠绕在它周围。</li>
	<li>创建一个 1/4 圆周截面，距离 PV 边缘约 0.5 mm（图 6A）。使用专用仪器（一根 27 G 针头，尖端切断并弯曲成 L 形）将盐水溶液通过管腔时扩大孔。</li>
	<li>使用直镊子握住袖带手柄并将其插入管腔。</li>
	<li>完全插入后，用 8-0 固定袖带丝线。单次结扎就足以进行固定。</li>
	<li>取下血管钳夹并重新灌注肝脏。</li>
	<li>小心地释放豌豆镊子并完成门静脉重建。整个光伏重建过程大约需要 5 分钟。</li>
</ol>5.肝下腔静脉重建
<ol><li>适当放置肝叶，并将连接到受体 IHIVC 背侧的 Pean 镊子移动到移植物的右下叶，用虎钳固定。</li>
	<li>用棉签轻轻擦拭，去除接受者 IVIHC 周围任何残留的肝组织。</li>
	<li>以8-0的比分丝线缠绕在接受者的 IVIHC 周围。</li>
	<li>在距受体的 IVIHC 边缘约 0.5 毫米处创建一个 1/4 圆周截面（图 6B）。</li>
	<li>将生理盐水溶液引入管腔后，将袖带插入管腔。 注意：由于颈部较短，袖带通常比 PV 中的插入得更浅。</li>
	<li>小心地用 8-0 固定袖带丝线，然后取下血管钳和豌豆钳。</li>
</ol>6.术后护理
<ol><li>利用加热垫和加热灯来促进术后恢复。</li>
	<li>持续监测血压、心率、呼吸频率、体温，以及食物和水的消耗。</li>
	<li>给药者皮下注射卡洛芬（5 mg/kg，每 6 小时或直到需要）以减轻术后疼痛。遵循当地的机构准则。 注意：如果小鼠的状况恶化，实验将立即停止，并通过吸入二氧化碳对小鼠实施安乐死。</li>
	<li>为了收集组织样本，重新麻醉小鼠并通过IACUC批准的方法对它们实施安乐死。</li>
</ol>

小鼠肝移植中有效血管吻合的策略

<ol>
	<li>Qian, S. G., Fung, J. J., Demetris, A. V., Ildstad, S. T., 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=Orthotopic+liver+transplantation+in+the+mouse.">Orthotopic liver transplantation in the mouse.</a> Transplantation. 52 (3), 562-564 (1991).</li><li>Yokota, 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=Orthotopic+mouse+liver+transplantation+to+study+liver+biology+and+allograft+tolerance.">Orthotopic mouse liver transplantation to study liver biology and allograft tolerance.</a> Nat Protoc. 11 (7), 1163-1174 (2016).</li><li>Shen, X. 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=Inflammatory+responses+in+a+new+mouse+model+of+prolonged+hepatic+cold+ischemia+followed+by+arterialized+orthotopic+liver+transplantation.">Inflammatory responses in a new mouse model of prolonged hepatic cold ischemia followed by arterialized orthotopic liver transplantation.</a> Liver Transpl. 11 (10), 1273-1281 (2005).</li><li>Kageyama, 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=Ischemia-reperfusion+injury+in+allogeneic+liver+transplantation:+A+role+of+CD4+T+cells+in+early+allograft+injury.">Ischemia-reperfusion injury in allogeneic liver transplantation: A role of CD4 T cells in early allograft injury.</a> Transplantation. 105 (9), 1989-1997 (2021).</li><li>Li, T., Hu, Z., Wang, L., Lv, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Details+determining+the+success+in+establishing+a+mouse+orthotopic+liver+transplantation+model.">Details determining the success in establishing a mouse orthotopic liver transplantation model.</a> World J Gastroenterol. 26 (27), 3889-3898 (2020).</li><li>Tian, Y., R&#252;diger, H. A., Jochum, W., Clavien, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+arterialized+and+nonarterialized+orthotopic+liver+transplantation+in+mice:+Prowess+or+relevant+model.">Comparison of arterialized and nonarterialized orthotopic liver transplantation in mice: Prowess or relevant model.</a> Transplantation. 74 (9), 1242-1246 (2002).</li><li>Steger, U., Sawitzki, B., Gassel, A. M., Gassel, H. J., Wood, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+hepatic+rearterialization+on+reperfusion+injury+and+outcome+after+mouse+liver+transplantation.">Impact of hepatic rearterialization on reperfusion injury and outcome after mouse liver transplantation.</a> Transplantation. 76 (2), 327-332 (2003).</li><li>Chen, 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=A+review+of+various+techniques+of+mouse+liver+transplantation.">A review of various techniques of mouse liver transplantation.</a> Transplant Proc. 45 (6), 2517-2521 (2013).</li><li>Kojima, 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=T+cell+carcinoembryonic+antigen-related+cell+adhesion+molecule+1-T+cell+immunoglobulin+domain+and+mucin+domain-containing+protein+3+cross+talk+alleviates+liver+transplant+injury+in+mice+and+humans.">T cell carcinoembryonic antigen-related cell adhesion molecule 1-T cell immunoglobulin domain and mucin domain-containing protein 3 cross talk alleviates liver transplant injury in mice and humans.</a> Gastroenterology. 165 (5), 1233-1248 (2023).</li><li>Yokota, S., Yoshida, O., Ono, Y., Geller, D. A., Thomson, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+the+mouse:+Insights+into+liver+immunobiology,+tissue+injury,+and+allograft+tolerance.">Liver transplantation in the mouse: Insights into liver immunobiology, tissue injury, and allograft tolerance.</a> Liver Transpl. 22 (4), 536-546 (2016).</li></ol>

任何作者都没有任何利益冲突需要披露。

学习血管重建是成功实现 MOLT 最具挑战性的方面。鉴于小鼠5 的体型较小，袖带的质量显着影响了重建的难度。本文提供了袖带准备、连接和重建的详细方案。
虽然与以前的报告在袖带准备和连接方面没有重大差异 2,5，但应考虑一些小问题。首先，袖带手柄的厚度应限制在总周长的 1/6 左右。如果太薄，手柄很容?...

小鼠原位肝移植（MOLT）是一种有效的实验方法，于1991年首次报道1。这种利用转基因小鼠和各种研究试剂的实验模型在研究冷暖缺血和再灌注损伤方面发挥了关键作用。然而，该模型的高技术复杂性阻碍了肝移植基础医学的发展2。MOLT 涉及三个主要步骤：（1） 从供体小鼠中取出肝脏，（2） 背台手术，以及 （3） 将肝脏植入受体。在这些受体手术中，血管吻合术构成了最大的挑战。虽然肝上下腔静脉吻合术通常通过手工缝合完成2，但肝下下腔静脉 （IHIVC） 和门静脉 （PV） 可以使用塑料袖带代替手工缝合线更有效地重建。
肝期表示切除受体的天然肝脏和移植物植入之间的间隔。为确保一致的结果，必须将肝脏分离时间限制在20分钟以内。因此，采用这一模式的研究仅限于特定机构3,4,5,6,7,8,9。在 MOLT 的各个阶段中，实现平稳的 PV 和 IHIVC 重建对于最大限度地减少肝功能时间并确保移植成功至关重要。
PV 和 IVC 重建通常使用血管袖带进行，因为与手工缝合线相比，袖带技术简化了血管吻合术 2,5,8。血管袖带准备和袖带安全连接所涉及的技术显着影响受体血管重建的复杂性。我们的目标是为与袖带方法相关的众多技术技巧提供详细的视觉指导，从而减少学习曲线。这些视频剪辑将清楚地了解如何将袖带连接到血管上，并在接受者手术期间重建 PV 和 IHIVC。

<table><tbody><tr><td>16 G intravenous catheter</td><td>TERUMO</td><td>SR-FF2032</td><td>IHIVC cuff</td></tr><tr><td>20 G intravenous catheter</td><td>TERUMO</td><td>SR-FF1651</td><td>PV cuff</td></tr><tr><td>8-0 braid silk</td><td>Natsume Seisakusho</td><td>CR9-80B2</td><td>8-0 silk</td></tr><tr><td>Belzer UW Cold Storage Solution&amp;nbsp;</td><td>Astellas</td><td></td><td>Organ preservation fluid</td></tr><tr><td>Bulldog clamp</td><td>B BRAUN</td><td>FB329R</td><td>Bulldog clamp</td></tr><tr><td>C57BL/6 mice&amp;nbsp;&amp;nbsp;</td><td>Oriental Bio Service</td><td></td><td></td></tr><tr><td>Isoflurane inhalation solution</td><td>Viatris</td><td></td><td>Anesthesic</td></tr><tr><td>Micro Blunted Tips 0.1 mm x 0.06 mm&amp;nbsp;</td><td>F.S.T</td><td>11253-20</td><td>Straight microforceps</td></tr><tr><td>Micro Serrefine Clamp Applicator with Lock</td><td>F.S.T</td><td>18056-14</td><td>Vessel clip applicator</td></tr><tr><td>Micro Serrefines&amp;nbsp;</td><td>F.S.T</td><td>18055-4</td><td>Vessel clip</td></tr><tr><td>No.11 Spare Blades</td><td>FEATHER Safety Razor</td><td>11</td><td>Blades</td></tr><tr><td>Ophthalmic scissor, round handle</td><td>B BRAUN</td><td>FD103R</td><td>Microscissor</td></tr><tr><td>Plastic rectangular-shaped container&amp;nbsp;&amp;nbsp;</td><td>Daiso</td><td></td><td>10 cm long, 15 cm wide and 6 cm high</td></tr><tr><td>SuperGrip Tips</td><td>F.S.T</td><td>00649-11&amp;nbsp;</td><td>Curved microforceps</td></tr><tr><td>SZX7</td><td>Olympus</td><td>SZX7</td><td>Microscope</td></tr><tr><td>Vise</td><td>Niigataseiki</td><td>00505351</td><td>A mechanical tool to secure Pean forceps on the table</td></tr></tbody></table>

vascular reconstruction with the cuff technique in mouse orthotopic liver transplantation

小鼠原位肝移植是研究肝缺血和再灌注损伤潜在机制的有效方法。然而，技术挑战对利用这种宝贵的实验模型并将这些技能传授给下一代构成了障碍。该手术最具挑战性的方面是血管重建，包括门静脉 （PV）、肝下下腔静脉 （IHIVC） 和肝上下腔静脉。使用塑料袖带而不是缝合线，可以更顺利地进行 PV 和 IHIVC 重建。通过将由静脉导管制成的袖带连接到移植物血管的尖端并将袖带插入受体血管来重建血管。两个最关键的方面是正确地观察血管的内腔和避免使用过度的力。我们的目标是提供在接受者手术中使用袖带技术进行血管重建的技术概述。这些袖带技术的技术技巧有望帮助显微外科医生促进血管重建并推进他们的研究。

当松开门静脉时，没有迂曲，肝脏均匀灌注时，PV 重建成功。肝功能时间应小于20分钟，因为肝脏时间超过25分钟会增加小鼠死亡的风险。如果移植物没有血液反流，则认为 IHIVC 重建成功。
使用器官保存溶液在低温下储存移植物1小时，导致再灌注后6小时的血清丙氨酸氨基转移酶水平约为2,000U / L（图7A）。移植后7天的存活率为100%（图7B</s...

Watch this Scientific Journal Video about Author Spotlight: Exploring Cold Ischemia and Warm Reperfusion Injury in Fatty Liver Transplantation at JoVE.com

作者聚焦：探索脂肪肝移植中的冷缺血和温再灌注损伤

该协议为在小鼠原位肝移植中使用袖带技术进行血管重建提供了技术信息。

袖带技术在小鼠原位肝移植中的血管重建

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. ...

我们的研究重点是肝移植，特别是冷缺血和温再灌注损伤。为此，我们使用小鼠肝移植模型作为研究工具。因为这项技术很困难，所以可能需要很长时间来学习和获得一致的结果。未来，我们想进行脂肪肝移植的研究。这仍然是临床肝移植需要克服的问题。

vascular-reconstruction-with-cuff-technique-mouse-orthotopic-liver

Research

JoVE Journal

Medicine

Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

1.2K 次观看.  Kyoto University. 我们的研究重点是肝移植，特别是冷缺血和温再灌注损伤。为此，我们使用小鼠肝移植模型作为研究工具。因为这项技术很困难，所以可能需要很长时间来学习和获得一致的结果。未来，我们想进行脂肪肝移植的研究。这仍然是临床肝移植需要克服的问题。

视频: 作者聚焦：探索脂肪肝移植中的冷缺血和温再灌注损伤

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

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 ...

科研

医学

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

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 ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

颈椎异位心脏移植小鼠

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

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 ...

Optimization of the Cuff Technique for Murine Heart Transplantation

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

穆林心脏移植袖口技术的优化

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to ...

免疫与感染

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

利用血管袖带的小鼠异位宫颈心脏移植

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

使用改性非缝合铐技术的宫颈主动脉移植模型

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...

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 ...

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

一种改进的袖带技术用于小鼠颈椎异位心脏移植模型

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...