Name: a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction
Uploaded: 2018-03-19T12:30:00.000+00:00
Duration: 14 min 33 s
Description: Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

这项工作得到教育部创新团队发展项目资助的支持 (no。IRT16R57), 中国国家自然科学基金 (81470896 号), 西安交通大学青年人才招聘计划研究基金 (cd-rw)。

该议定书是按照《护理和使用实验动物准则》进行的, 并获得了西安交通大学动物实验伦理学委员会的批准。
注意: 本协议中的任何程序都没有在手术显微镜下进行。
1. 术前准备
<ol><li>磁环的设计和形状 (图 1)<ol>
<li>通过匹配一对钕铁硼磁环来组装磁吻合装置。</li>
		<li>根据大直径、小轴直径、厚度和重量8毫米、5.5 毫米、1毫米、0.5 克等参数, 设计 SHVC 的环。</li>
		<li>用线电极切割技术将环按所需形状曲线, 然后用钛氧化物喷枪将所获得的环涂上, 用500毫克/升乙烯氧化物在55-60 摄氏度上消毒6小时。</li>
	</ol>
</li>
	<li>袖口 (图 2)<ol>
<li>通过切割聚乙烯管, 准备 IHVC 和 PV 吻合的袖口。确保袖口的身体是5毫米长, 2 毫米袖口延长。 注: PV 袖口的内外直径为1.8 和2.1 毫米, 而 IHVC 袖口的内径和外径分别为2.6 和2.8 毫米。</li>
	</ol>
</li>
	<li>支架 (图 2)<ol>
<li>通过切割24克长5毫米的静脉插管, 在两端产生倾斜, 准备用于吻合的支架。</li>
	</ol>
</li>
	<li>实验动物<ol>
<li>使用雄性大大鼠体重在250和 280 g 之间作为捐助者和受援国。确保接收人的体重与捐献者的重量相同或稍重 (< 10 克)。</li>
		<li>把老鼠留在气候控制的房间里, 在手术前可以免费获得食物和水。</li>
	</ol>
</li>
</ol>2. 捐助者的行动
<ol><li>术前1小时注射丁丙诺啡 (0.05 毫克/千克) 止痛。</li>
	<li>异氟醚吸入麻醉供体鼠。使用5体积% 异氟醚, 在1.5 升/分钟的一个有机玻璃盒中的流量, 用于诱导麻醉和2体积% 异氟醚的流速 0.6-0.8 升/分钟在一个锥面具, 以维持麻醉。通过执行脚趾捏和皮肤捏来确认麻醉深度。</li>
	<li>使用电动剃须刀剃去老鼠的整个腹部皮肤, 以允许更清洁的暴露。用聚维酮碘溶液对相应皮肤进行消毒。</li>
	<li>从从纵切口中点延伸到 midaxillary 线的1厘米以上的 xiphisternum 沿linea, 通过十字切口打开腹腔。将胃肠道向左 Exteriorize, 用湿纱布盖住。</li>
	<li>用止血钳将 xiphisternum 向头部拉紧。解剖镰韧带和肝周围的结缔组织。用6-0 丝缝线分离和结扎左下膈静脉。</li>
	<li>将支架插入胆管。<ol>
<li>将 BD 钳在胃十二指肠静脉加入门静脉的点上方, 并将其拉紧, 用 microforceps 维持少许张力。然后执行一个 "V" 切口1毫米长度在前壁约5毫米近到胆道汇合由一个微型剪刀。</li>
		<li>用弯曲的 microforceps 将支架插入胆管腔内。确保至少有一半的支架位于胆管外。</li>
		<li>用6-0 丝缝线固定支架。切开缝合的一端。在以后的吻合过程中保持另一条。在支架下面横断面胆管, 确保胆汁能从支架中出来。</li>
	</ol>
</li>
	<li>将 IHVC 分离到左肾静脉水平。用棉花芽夹层从前后周围组织中释放 IHVC, 直到 IHVC 骷髅。用6-0 丝缝线分离和结扎右肾上腺静脉和右肾静脉。</li>
	<li>Heparinize 注射 50 U 肝素稀释2毫升的正常盐水溶液通过该鸡的背静脉。</li>
	<li>灌注肝脏。<ol>
<li>将腹主动脉隔离在左肾静脉下方。将22克导管插入主动脉。</li>
		<li>执行开胸术, 用 microforceps 钳住胸主动脉。</li>
		<li>灌注通过导管在主动脉中, 用20毫升的生理盐水溶液, 含有 25 U 肝素 (4 °c), 以3毫升/分钟的速度与输液泵。同时, 在右肾静脉下切开腔静脉, 使灌注液流出肝脏。</li>
		<li>当供肝变苍白时, 将 SHVC 与部分隔膜和 IHVC 在左肾静脉的水平上进行横断面。</li>
	</ol>
</li>
	<li>用两个微钳解剖 PV 周围的结缔组织。结扎和分裂的幽门静脉与8-0 普理灵缝合。将 PV 在脾脏静脉的水平上进行样带。将供肝切除, 用冷生理盐水冲洗。安置它在一个冷的盐水浴4°c。</li>
</ol>3. 接枝制剂
注: 所有肝移植制剂的手术步骤均在4摄氏度的冷盐水浴中进行。
<ol><li>将袖口附加到捐赠者的 IHVC (图 3)。<ol>
<li>将 IHVC 插入袖口, 袖伸指向肝脏。将袖伸放在 IHVC 后壁上。修复袖口扩展和 IHVC 与弯曲 microforceps 本身附加使用文件剪辑到浴缸容器。</li>
		<li>用两个微钳将 IHVC 的远端埃弗特在袖体上。</li>
		<li>用6-0 丝缝线保护边缘。确保在过程中 IHVC 不扭曲。</li>
	</ol>
</li>
	<li>重复步骤3.1 在 pv 上附加一个袖口的捐助者的 pv。</li>
	<li>将磁环连接到捐赠者的 SHVC (图 4)。<ol>
<li>拉隔膜通过四钳和修剪 SHVC。</li>
		<li>采用钛合金微钳, 通过磁环 SHVC。</li>
		<li>埃弗特在环上 SHVC 的远端。</li>
		<li>用6-0 丝缝线把戒指固定起来。确保在过程中 SHVC 不扭曲。</li>
		<li>清除肝脏周围的多余隔膜。</li>
	</ol>
</li>
	<li>将供肝浸入冷生理盐水浴中, 贮存在摄氏4摄氏度。</li>
</ol>4. 收件人操作
注意: 接收鼠移植植入的模式显示在图 5中。
<ol><li>术前1小时注射丁丙诺啡 (0.05 毫克/千克) 止痛。异氟醚吸入麻醉供体鼠。使用5体积% 异氟醚, 在1.5 升/分钟的一个有机玻璃盒中的流量, 用于诱导麻醉和2体积% 异氟醚的流速 0.6-0.8 升/分钟在一个锥面具, 以维持麻醉。</li>
	<li>剃掉老鼠的整个腹部皮肤。用聚维酮碘溶液对相应皮肤进行消毒。</li>
	<li>通过4厘米中线切口打开腹腔。把腹部拉钩。把肋骨从中线尽可能地分开。</li>
	<li>用止血钳将 xiphisternum 向头部拉紧。解剖肝脏周围的结缔组织和韧带。结扎和分裂肝-食管韧带。结扎和分肝动脉8-0 普理灵缝合。</li>
	<li>从第一个肝门处分离出胆总管。将6-0 丝缝合周围的胆总管在其分裂和结扎的普通胆管。</li>
	<li>解剖 IHVC 到右肾静脉。结扎和分裂右肾上腺静脉。在肝脏的 SHVC 上放一条带子。</li>
	<li>用微血管夹子钳住右肾静脉的 IHVC。在幽门静脉的水平上用微血管夹钳住 PV。<ol>
<li>将血液从肝脏中冲洗出来, 慢慢地通过 PV 注入2毫升生理盐水。拉下原来放置的皮带, 拖下肝脏。用 Satinsky 钳夹住 SHVC 和部分隔膜。</li>
		<li>横断面的 IHVC 接近的实质和 PV 在门肝。transecting 肝上方的 SHVC, 将肝脏切除。快速取出受体肝。</li>
	</ol>
</li>
	<li>将磁环连接到接收者的 SHVC。<ol>
<li>将收件人剩余的 SHVC 插入磁环中。</li>
		<li>用磁吸引固定 Satinsky 钳上的磁环。</li>
		<li>埃弗特 SHVC 在磁环上, 直到容器覆盖环的边缘。</li>
	</ol>
</li>
	<li>将供肝移植原位, 用冷湿纱布盖住。</li>
	<li>用正常的盐水溶液填充捐献者的腔体和受体的 SHVCs。彻底清除气泡以防止空气栓塞。通过磁力耦合在供体和接收器 SHVC 中嵌入的磁环, 完成 SHVC 重建。</li>
	<li>重建光伏。<ol>
<li>通过拉动8-0 普理灵缝合以保持 pv 的张力, 对接收者的 pv 的右、左分支进行插拔。</li>
		<li>将接受者的 PV 钳从幽门静脉移至脾静脉, 以充分暴露静脉。</li>
		<li>切开接受者的前壁。用正常的盐水溶液填充 PV 的流明。</li>
		<li>用弯曲钳握住捐赠者 PV 的袖口。</li>
		<li>将捐赠者的 pv 的袖口体插入接收者的 pv, 同时保持正常生理盐水冲洗 pv 腔。</li>
		<li>用圆周6-0 的丝结扎保护它。通过在 PV 和 SHVC 上释放钳位, 重新建立移植肝的血流。</li>
	</ol>
</li>
	<li>在 IHVC 上重复步骤4.11 以重建 IHVC。</li>
	<li>用支架技术重建胆管。<ol>
<li>在接受者胆总管前壁切开 "V"。</li>
		<li>将供体支架插入受体 BD 的腔内, 并用6-0 丝环缝固定。将两个胆管的缝线拉在一起, 并系上缝合线, 使 BD 靠近对方。</li>
		<li>包裹在吻合部位的网膜组织, 以防止胆漏。</li>
	</ol>
</li>
	<li>用温热生理盐水溶液冲洗腹腔。用连续3-0 个不可吸收的手术缝合在两层闭合腹部切口。</li>
	<li>术后立即皮下注射16毫克/千克头孢呋辛 (抗生素) 和0.05 毫克/千克丁丙诺啡 (止痛药)。</li>
	<li>把老鼠放在干净的笼子里, 让食物和水广告随意。</li>
	<li>每12小时向腹腔注射头孢呋辛 (16 毫克/千克) 和丁丙诺啡 (0.05 毫克/千克) 3 天。</li>
</ol>

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作者没有什么可透露的。

许多临床进展在肝脏移植可以归因于动物研究。大鼠肝移植是在器官保存、移植免疫学、生理学和病理学研究中广泛应用和公认的模型。然而, 这也是一个非常复杂的过程, 需要先进的显微外科技术。尽管在大鼠肝移植手术过程中有许多改进, SHVC 吻合仍然是一个具有挑战性的步骤。本文介绍了一种新的大鼠肝移植 SHVC 重建的磁性吻合技术。该技术的应用减少了 SHVC 重建到2分钟以下的时间, 并将大?...

大鼠原位肝移植模型是移植研究的关键1,2。在1973年的3中, 第一个老鼠肝移植是由李et . 描述的。在该模型中, 所有的血管都是用手缝技术重建的。手工缝合技术需要先进的显微外科技术, 这大大限制了它的使用。从那时起, 对原《大鼠肝移植术》的各种修改进行了报道。其中, 1979年卡玛达et. 报告的门静脉 (PV) 和 infrahepatic 腔静脉 (IHVC) 吻合的袖套技术被认为是这一模型的一个重大改进, 因为它大大简化了重建程序4.然而, 由于接受者的 suprahepatic 腔静脉 (SHVC) 的前壁短, 袖套技术很难用于重建 SHVC。该领域的大多数研究人员仍然使用手工缝合技术进行 SHVC 重建, 这使得它成为大鼠肝移植的瓶颈步骤5,6,7。
磁吻合技术 (即, magnamosis) 是一种连接两个容器或其他管状结构的方法, 使用两个磁体8、9、10、11之间的吸引力。磁力逐渐压缩和改造组织成一个强有力的, 无缝线切口吻合12, 13.这种压缩吻合已被证明是有效的人14,15。我们设计了一双磁性环, 专门用于 SHVC 的大鼠的吻合。我们最近的研究表明, 磁吻合技术优于手缝合技术的 SHVC 重建的大鼠肝移植术16。本文的目的是提供一个详细的, 循序渐进的协议, 为 SHVC 重建的大鼠使用新的磁性吻合技术。

<table><tbody><tr><td>Anesthesia Machine</td><td>Harvard</td><td>&amp;nbsp;tabletop</td><td>Animal anaesthesia</td></tr><tr><td>PLX7000B HF Mobile Digital C-arm System</td><td>Perlong Medical</td><td>PLX7000B</td><td>It is mainly used for the angiography and photography of various operations</td></tr><tr><td>Syringe Pump</td><td>Mindray</td><td>BeneFusion SP5</td><td>intravenous infusion</td></tr><tr><td>Isoflurane</td><td>RWD life Science Co.</td><td></td><td>anesthetic:for the induction and maintenanceof anesthesia</td></tr><tr><td>iohexol</td><td>Shanghai General Pharmaceutical Co</td><td></td><td>&amp;nbsp;intravascular contrast media&amp;nbsp;&amp;nbsp;</td></tr><tr><td>heparin sodium injection</td><td>SPH No.1 Biochemical &amp;amp; Pharmaceutical Co., LTD&amp;nbsp;</td><td></td><td>&amp;nbsp;prevent the formation of thrombosis&amp;nbsp;</td></tr><tr><td>cefuroxime&amp;nbsp;</td><td>Glaxo Operations UK Limited&amp;nbsp;</td><td></td><td>an antibiotic</td></tr><tr><td>buprenorphine&amp;nbsp;</td><td>TIPR Pharmaceutical Responsible Co.,Ltd</td><td></td><td>an analgesic</td></tr><tr><td>curved microforceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>W40350</td><td>surgical tool</td></tr><tr><td>hemostatic forceps(straight)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J31010</td><td>surgical tool</td></tr><tr><td>hemostatic forceps(curved)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J31020</td><td>surgical tool</td></tr><tr><td>Satinsky clamp</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>XEC050</td><td>surgical tool</td></tr><tr><td>needle holder</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J32010</td><td>surgical tool</td></tr><tr><td>microneedle holder</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WBA040</td><td>surgical tool</td></tr><tr><td>notched forceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J42010</td><td>surgical tool</td></tr><tr><td>tissue forceps(with hook)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>J41010</td><td>surgical tool</td></tr><tr><td>tissue scissor</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>Y00040</td><td>surgical tool</td></tr><tr><td>surgical scissors</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>Y00030</td><td>surgical tool</td></tr><tr><td>micro scissors</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>MR-S121T</td><td>surgical tool</td></tr><tr><td>microvessel clips</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>XEC240</td><td>surgical tool</td></tr><tr><td>straight microforceps(titanium alloy)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WCC010</td><td>surgical tool</td></tr><tr><td>curved microforceps&amp;nbsp; (titanium alloy)</td><td>Shanghai Medical Instruments (Group) Ltd., Corp.</td><td>WCC020</td><td>surgical tool</td></tr></tbody></table>

a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction

大鼠原位肝移植模型是移植研究的基础。这是一个非常复杂的动物模型, 需要一个陡峭的学习曲线。采用袖套技术对门静脉 (PV) 和 infrahepatic 腔静脉 (IHVC) 进行吻合, 大大简化了大鼠移植过程。然而, 由于接受者的 suprahepatic 腔静脉 (SHVC) 的前壁短, 袖套技术很难用于重建 SHVC。该领域的大多数研究人员仍然使用手缝合技术进行 SHVC 重建, 这使得它成为大鼠原位肝移植的瓶颈步骤。磁吻合技术 (即, magnamosis) 是一种用两个磁体之间的引力来连接两个容器的方法。我们最近的研究表明, 磁吻合技术优于手缝合技术的 SHVC 重建的大鼠。在本文中, 我们展示了一个逐步的协议, SHVC 重建的大鼠使用新的磁性吻合技术。在该模型中, 采用标准袖套技术对 PV 和 IHVC 进行重建, 采用支架技术进行胆管重建。肝脏再动脉没有执行。磁吻合技术使 SHVC 重建更容易, 大大缩短了 anphepatic 期。经过合理的学习曲线, 即使没有先进的显微外科技术的研究人员也能用这种大鼠肝移植模型产生可靠和可重复的结果。

经过大约10次尝试, SHVC 重建的磁吻合技术成功地掌握在一个没有事先进行显微外科训练的研究员身上。SHVC 的重建花费不到2分钟。无肝期阶段为接受者老鼠大约10分钟。再灌注后1、5、30天 SHVC 吻合口无血栓、出血或 angiostegnosis。下腔 cavography 在移植后2周进行。如图 6所示, 磁环完好无损, 血液流动是通过 SHVC 吻合的专利。移植后存活率为95% 天 1, 90% 天...

Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

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

suprahepatic 腔静脉 (SHVC) 的重建仍是大鼠原位肝移植的难点。本文介绍了一种新型的磁吻合技术在大鼠 SHVC 重建中的应用。

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 ...

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JoVE Journal

Medicine

9.8K 次观看.  First Affiliated Hospital, Xi'an Jiaotong University. suprahepatic 腔静脉 (SHVC) 的重建仍是大鼠原位肝移植的难点。本文介绍了一种新型的磁吻合技术在大鼠 SHVC 重建中的应用。

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

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

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 ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

大鼠异位心脏腹部/在卷装配置单肺移植

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

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. ...

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 ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

无静脉-静脉旁路术的猪肝移植作为扩展标准供体模型

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...

小鼠肝移植和同种异体移植物通畅性分析

Successful Orthotopic Liver Transplantation in Mice Utilizing Microcomputed Tomography Angiography

Microcomputed tomography (microCT) angiography is an invaluable resource to researchers. New advances in this technology have allowed for high-quality images to be obtained of micro-vasculature and are high-fidelity tools in the field of organ transplantation. In this model of orthotopic liver transplantation (OLT) in mice, microCT affords the opportunity to evaluate allograft anastomosis in real time and has the added benefit of not having to sacrifice study animals. The choice of contrast, as well as image acquisition settings, create a high-definition image, which gives researchers invaluable information. This allows for evaluation of the technical aspects of the procedure as well as potentially evaluating different therapeutics over an extended duration of time. In this protocol, we detail an OLT model in mice in a stepwise fashion and finally describe a microCT protocol that can give high-quality images, which aid researchers in in-depth analysis of solid organ transplantation. We provide a step-by-step guide for liver transplantation in a mouse, as well as briefly discuss a protocol for evaluating the patency of the graft through microCT angiography.

作者聚焦:通过小鼠模型中的 MicroCT 血管造影增强移植研究 

In this protocol, we discuss the implementation of a model of successful orthotopic liver transplantation (OLT) in mice. Additionally, adjuvants to further analyze allograft patency after successful OLT in a mouse are discussed as well, specifically utilizing microcomputed tomography (microCT) scans.

利用显微计算机断层扫描血管造影成功对小鼠进行原位肝移植

Microcomputed tomography (microCT) angiography is an invaluable resource to researchers. New advances in this technology have allowed for high-quality ...

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

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此视频中的章节

大鼠原位肝移植

部分原位肝移植大鼠模型的肝动脉重建的外科手术

在急性肝衰竭异位辅助性大鼠肝移植流量调节门静脉动脉

大鼠异位心脏腹部/在卷装配置单肺移植

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

矫形肝移植大鼠模型中动脉重联后并发症减少

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

无静脉-静脉旁路术的猪肝移植作为扩展标准供体模型

利用显微计算机断层扫描血管造影成功对小鼠进行原位肝移植

利用显微计算机断层扫描血管造影成功对小鼠进行原位肝移植