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, 并将大鼠肝移植的无肝期期缩短到约10分钟。我们认为这种技术在移植研究中具有重要的意义。
使用这种磁吻合技术重建 SHVC 的关键步骤是将磁环连接到捐献者和接收者的 SHVC。每次小心都应该被用来避免扭曲 SHVC。此外, 在连接两个磁环时, SHVC 中的气泡应彻底去除。
使用此技术有一些限制。Fischer-344 大鼠和水牛大鼠等几株大鼠的 SHVC 非常短。将磁环连接到它们的 SHVC 是不可能的。因此, 它们不适合这种技术。然而, 该技术可以很容易地应用于大多数品系的大鼠肝移植, 包括大大鼠、路易斯大鼠和棕色挪威大鼠。磁环可以干扰磁共振成像 (MRI) 扫描。因此, 如果实验需要核磁共振检查, 就不能使用这种磁吻合技术。
总之, 磁吻合技术使大鼠容易快速 SHVC 重建成为可能。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。

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。

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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 3, 85% 天7至 30 (图 7)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56933/56933fig1.jpg"> 图 1:用于吻合 SHVC 的磁环的设计 (A) 和照片 (B)。 SHVC: suprahepatic 腔静脉。 <a href="/files/ftp_upload/56933/56933fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56933/56933fig2.jpg"> 图 2:IHVC (A)、PV (B) 和 BD (C) 的吻合的袖口和支架。 IHVC: infrahepatic 腔静脉;光伏: 门静脉;胆管。<a href="/files/ftp_upload/56933/56933fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56933/56933fig3.jpg"> 图 3:IHVC 和 PV 的袖套吻合模式。将 IHVC 或 PV 插入袖口 (a);使用两个 microforceps (B) 将 IHVC 的远端或 PV 埃弗特在袖口体上;用6-0 丝缝线固定边缘 (C);将捐赠者的 IHVC 或 pv 的袖口体插入收件人的 IHVC 或 pv (D)。 IHVC: infrahepatic 腔静脉;PV: 门静脉。<a href="/files/ftp_upload/56933/56933fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56933/56933fig4.jpg"> 图 4:SHVC 的磁性吻合模式。使用钛合金 microforceps (a) 通过磁环 SHVC;埃弗特环上 SHVC 的远端 (B);用6-0 丝缝线固定环 (C)。通过磁力 (D) 将在捐赠者和接收者的 SHVC 中嵌入的磁环耦合在一起。 SHVC: suprahepatic 腔静脉。<a href="/files/ftp_upload/56933/56933fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56933/56933fig5.jpg"> 图 5:受体大鼠移植植入的模式。SHVC 的重建: 磁性吻合术;IHVC 与 PV 的重建: 袖套吻合术;重建的 BD: 支架吻合术。重建 HA: 未执行。 SHVC: suprahepatic 腔静脉;IHVC: infrahepatic 腔静脉;光伏: 门静脉;胆管: 胆道;肝动脉。<a href="/files/ftp_upload/56933/56933fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56933/56933fig6.jpg"> 图 6:移植后2周大鼠下腔 cavography。<a href="/files/ftp_upload/56933/56933fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56933/56933fig7.jpg"> 图 7:有代表性的生存治疗后移植。<a href="/files/ftp_upload/56933/56933fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

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 ...

a-rat-model-orthotopic-liver-transplantation-using-novel-magnetic

Nanyang Technological University

Research

JoVE Journal

Medicine

9.7K 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 Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

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

Electrolytic Inferior Vena Cava Model (EIM) of Venous Thrombosis

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential therapeutic compounds (1). New compounds to be utilized in the treatment and prevention of DVT are currently being developed. The delivery of potential therapeutic antagonist compounds to an affected thrombosed vein has been problematic. In the context of therapeutic applications, a model that uses partial stasis and consistently generates thrombi within a major vein has been recently established. The Electrolytic Inferior vena cava Model (EIM) is mouse model of DVT that permits thrombus formation in the presence of continuous blood flow. This model allows therapeutic agents to be in contact with the thrombus in a dynamic fashion, and is more sensitive than other models of DVT (1). In addition, this thrombosis model closely simulates clinical situations of thrombus formation and is ideal to study venous endothelial cell activation, leukocyte migration, venous thrombogenesis, and to test therapeutic applications (1). The EIM model is technically simple, easily reproducible, creates consistent thrombi sizes and allows for a large sample (i.e. thrombus and vein wall) which is required for analytical purposes.

Electrolytic Inferior Vena Cava Model EIM of Venous Thrombosis

The electrolytic induction of endothelial activation to the internal surface of the Inferior Vena Cava results in venous type thrombus formation due to endothelial activation and partial blood stasis, two components of Virchow's triad.

电解下腔静脉静脉血栓形成的模型（EIM）

Animal models serve a vital role in deep venous thrombosis (DVT) research in order to study thrombus formation, thrombus resolution and to test potential ...

Mouse Complete Stasis Model of Inferior Vena Cava Thrombosis

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE). In the United States (U.S.), the high morbidity and mortality rates make VTE a serious health concern 1-2. After heart disease and stroke, VTE is the third most common vascular disease 3. In the U.S. alone, there is an estimated 900,000 people affected each year, with 300,000 deaths occurring annually 3. A reliable in vivo animal model to study the mechanisms of this disease is necessary.
The advantages of using the mouse complete stasis model of inferior vena cava thrombosis are several. The mouse model allows for the administration of very small volumes of limited availability test agents, reducing costs dramatically. Most promising is the potential for mice with gene knockouts that allow specific inflammatory and coagulation factor functions to be delineated. Current molecular assays allow for the quantitation of vein wall, thrombus, whole blood, and plasma for assays. However, a major concern involving this model is the operative size constraints and the friability of the vessels. Also, due to the small IVC sample weight (mean 0.005 grams) it is necessary to increase animal numbers for accurate statistical analysis for tissue, thrombus, and blood assays such as real-time polymerase chain reaction (RT-PCR), western blot, enzyme-linked immunosorbent (ELISA), zymography, vein wall and thrombus cellular analysis, and whole blood and plasma assays 4-8.
The major disadvantage with the stasis model is that the lack of blood flow inhibits the maximal effect of administered systemic therapeutic agents on the thrombus and vein wall.

The mouse complete stasis model of inferior vena cava thrombosis yields quantifiable amounts of vein wall tissue and thrombus. It has proven useful for evaluating interactions between the vein wall and the occlusive thrombus and in assessing the progression from acute to chronic inflammation.

鼠标完成血瘀模型的下腔静脉血栓

Venous thromboembolism (VTE) includes both deep vein thrombosis (DVT) and pulmonary embolism (PE).  In the United States (U.S.), the high morbidity and ...

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 ...

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 ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

下腔静脉间置移植物在小鼠模型中植入

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

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 ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Patch Angioplasty in the Rat Aorta or Inferior Vena Cava

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, we developed a rat model of patch angioplasty that can be used in either a vein or an artery, creating a patch venoplasty or a patch arterioplasty, respectively. Technical aspects of this model are discussed. The infra-renal IVC or aorta are dissected and then clamped proximally and distally. A 3 mm venotomy or arteriotomy is performed in the infrarenal inferior vena cava or aorta of 6 to 8 week-old Wistar rats. A bovine pericardial patch (3 mm x 1.5 mm x 0.6 mm) is then used to close the site using a 10-0 nylon suture. Compared to arterial patches, venous patches show increased neointimal thickness on postoperative day 7. This novel model of pericardial patch angioplasty can be used to examine neointimal hyperplasia on vascular biomaterials, as well as to compare the differences between the arterial and venous environments.

We have established a model of pericardial patch angioplasty that can be used in either small-diameter veins or arteries. This model can be used to compare venous and arterial neointimal hyperplasia formation.

血管补片在大鼠主动脉和下腔静脉

Pericardial patches are commonly used in vascular surgery to close vessels. To facilitate studies of the neointimal hyperplasia that forms on the patch, ...

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 ...