Name: re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy
Uploaded: 2018-04-08T13:00:00.000+00:00
Duration: 14 min 52 s
Description: Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

本研究获得中国国家自然科学基金 (81501382 号赠款) 和浙江省自然科学基金 (批准号:) 的支持。LQ13H100003 和 LQ16H100002)。

所有的程序都遵循了温州医科大学动物政策和福利委员会批准的《啮齿动物外科指南》 15。
1. 动物
<ol><li>用雄性 Lewis 老鼠称250-350 克作为捐献者和受体。</li>
</ol>2. 操作环境
<ol><li>在手术显微镜下执行所有显微外科手术, 放大率从7X 到20X 不等。手术前要消毒所有外科器械。 注意: 操作室和操作表清洁, 但不消毒。</li>
</ol>3. 基本显微手术机动
<ol><li>自由, 解剖, 并动员船只。<ol>
<li>首先, 使用直的微钳来解除血管周围的组织。</li>
		<li>第二, 反复轻轻地打开和关闭弯曲微钳的尖端, 这样细微的提示就可以切开并剥开血管前壁上的周围组织。</li>
		<li>第三, 使用直式微钳钳夹并提起血管以暴露后间隙。然后, 使用弯曲微钳的尖端剥离血管后壁上的周围组织。最后, 一个没有任何周围组织连接的秃头血管应该被提出。</li>
	</ol>
</li>
	<li>分裂或双结扎, 并划分血管或组织。<ol>
<li>首先, 使用两个微钳16在容器或组织上系两个方结。第二, 用微剪刀切断两个方结之间的血管或组织。</li>
	</ol>
</li>
</ol>4. 袖套和胆道支架的制备
<ol><li>用12口径和14口径静脉导管分别制作门静脉和 infrahepatic 腔静脉的袖口, 而胆道支架则采用22口径静脉导管。袖口由身体部分和手柄部分组成。使用身体部位的沉, 将容器壁固定在袖口上 (图 1)。</li>
</ol>5. 大鼠的麻醉和固定
<ol><li>在5% 异氟醚与0.5 升/敏氧混合的感应箱中轻轻放置路易斯大鼠, 诱导吸入麻醉。监测大鼠的呼吸频率, 以防止过量异氟醚引起的意外死亡。减少异氟醚的浓度为 2%, 以保持麻醉时, 大鼠不响应疼痛刺激。</li>
	<li>将老鼠移动到一个30厘米 x 30 厘米的橡木板上, 用消毒的手术垫毛巾盖住, 然后把老鼠放在仰卧位。用内侧胶带绑住大鼠的四只四肢, 随后用四图钉固定胶带。一次性用70% 酒精消毒皮肤, 碘伏三次。</li>
</ol>6. 捐助者的行动
<ol><li>暴露手术现场<ol>
<li>在上腹部切开一小横切口, 用电动凝固将两侧下腹血管横断面。</li>
		<li>用手术剪刀将横切口伸展至上腹部的肋缘。</li>
		<li>反向和固定上腹壁和剑突的过程, 分别对颅侧。</li>
		<li>用两根湿棉布把肠子从腹腔移出。用一块湿纱布盖住肠道。</li>
	</ol></li>
	<li>切断韧带和修剪 suprahepatic 腔静脉 (SHVC)<ol>
<li>falciformal 韧带、冠状动脉韧带和三角韧带依次切除。</li>
		<li>用微钳从隔膜中释放左膈静脉。然后, 双结扎使用6-0 丝缝线。 注意: 不要在这个步骤中划分这个静脉。</li>
		<li>用微剪刀切断 gastrohepatic 韧带, 释放下尾叶和 hepatoesophageal 韧带和动脉。</li>
	</ol></li>
	<li>在切除前阻断微光和 ML 的血液供应 (图 2)<ol>
<li>首先, 使用弯曲钳仔细解剖的公共树干的后壁, 提供微光和左中位叶 (爱我的生活) 从实质。第二, 双结扎, 并将其与相应的胆管和肝动脉分开。</li>
		<li>轻轻地解剖和分割 RML 的 PV, 因为这种静脉紧贴肝脏实质。 注: 建议使用14X 放大倍数。不要伤害右上叶的 pv, 这是接近 RML 的 pv。</li>
		<li>鉴别肝动脉和胆管的 RML 位于 PVs 之间的 RML 和爱我的生活。双结扎, 用6-0 丝缝线将 RML 的肝动脉和胆管分开。 注意: 从红色到深红色的微光和 ML 颜色的变化是成功阻断血液供应的指标。深入了解血管解剖是识别和解剖相应血管的必要条件。以前的研究在大鼠清楚地说明了肝脏解剖由腐蚀铸造10。</li>
	</ol></li>
	<li>切除微光<ol>
<li>结扎的椎弓根使用3-0 周丝缝合咬合左外侧肝静脉的血流。使用微剪刀, 切断肝脏立即结扎以上切除肝脏肿块。</li>
	</ol></li>
	<li>用我的生命去切除爱<ol>
<li>用蚊子止血钳夹住中位裂 (图 3A) 中的左中位肝静脉。用微剪刀把爱我生命中的所有肝实质都清除掉。</li>
		<li>在左中位肝静脉周围的镊子下, 放置一条穿透性缝合线, 立即渗入肝实质。然后, 在松开镊子之前, 系上一个方形结结扎。</li>
		<li>在镊子下再放两个穿刺缝线, 环绕着我生命中敞开的伤口。然后, 用两个方结来缝合切口的其余部分。释放蚊子止血钳, 检查是否有出血。</li>
	</ol></li>
	<li>正在切除 RML (图 4)<ol>
<li>放置蚊子止血钳, 将 RML 的底座环绕在0.5 厘米至 SHVC 的距离 (图 4B)。然后, 用蚊子止血钳交叉夹住底座, 咬合右中肝静脉。</li>
		<li>使用微剪刀, 去除所有的肝实质的 RML 沿上表面的镊子。一定要非常接近钳面, 留下一个普通的和薄残端的 ML。</li>
		<li>将第一条穿刺缝线穿透在镊子下的肝实质包围中位肝静脉。然后, 用方结结扎中位肝静脉。用同一方法将第二穿刺缝合结扎右中肝静脉。在6.5.2 的描述之后, 再放置两条穿孔缝合线, 靠近切口的其余部分。</li>
	</ol></li>
	<li>infrahepatic 腔静脉的修整 (IHVC)<ol>
<li>切开打开的腹腔, 露出 IHVC。用弯曲的微钳从周围组织中释放 IHVC 和右肾静脉。解剖和结扎右肾上腺静脉, 从后方排泄到 IHVC。</li>
		<li>在结扎右肾静脉和右肾动脉后, 进行正确的肾切除术, 以节省足够的血管长度, 以便在以后的步骤中将袖口安装到 IHVC 上。</li>
	</ol></li>
	<li>安装胆道支架<ol>
<li>用微钳从第一个肝门动员胆总管 (CBD)。结扎在幽门静脉的交叉点和 cbd 使用6-0 丝缝合。</li>
		<li>切开前壁的一个横切口在 CBD 的近侧结扎。插入一个由22口径静脉注射导管组成的胆道支架, 通过切口进入 CBD 的腔内。用圆周6-0 丝缝线固定胆道支架。在6-0 丝缝线之间进行样条。</li>
	</ol></li>
	<li>修整光伏<ol>
<li>结扎的幽门静脉使用6-0 丝缝合远端的 PV。接下来, 结扎的幽门静脉使用7-0 聚丙烯缝合接近 PV。在这两个结扎之间的幽门静脉样条。双结扎, 并以同样的方式划分脾静脉。 注: 用7-0 聚丙烯缝线结扎血管, 使其与6-0 丝缝线相比, 更小的结扎结。一个大的结可能会干扰袖口安装在最后一步。幽门静脉和脾静脉的血管壁薄, 因此, 在解剖过程中, 温和的操作是必不可少的。在17手术前, 应阅读一篇以前发表的文章, 其中有更多关于解剖和划分幽门静脉和脾静脉的技术说明和插图。</li>
	</ol></li>
	<li>普通肝动脉的修整 (茶)<ol>
<li>用两个微钳从周围组织中仔细地释放茶及其两个分岔、GDA 和胃十二指肠动脉。双结扎的 GDA 使用6-0 丝缝线在岔 GDA。</li>
	</ol></li>
	<li>Heparinizing 与灌注肝移植术<ol>
<li>在1毫升生理盐水中通过小弟弟静脉注入50的肝素。等待5分钟, 实现系统性肝素化。</li>
		<li>用三弯曲微 serrefines 对 IHVC 和茶进行交叉钳咬合肝血流。在 PV 前壁上做一个小切口, 插入22口径导管。灌注在 20 cmH2O 的压力下, 通过导管在4摄氏度的部分肝移植。</li>
		<li>用手术剪刀快速切开胸腔和胸腔静脉, 排出血液和灌流液。不要停止灌注, 直到整个部分肝移植的颜色均匀变黄。</li>
	</ol></li>
	<li>移植部分肝移植术<ol>
<li>用微剪刀对两个结扎 (6.2.2 中描述的) 左膈静脉进行样条。</li>
		<li>接下来, 横断面 SHVC 靠近隔膜, 以保持血管长度尽可能长。用微剪刀在脾脏静脉的水平进行样带。将两个结扎之间的 GDA, 并与腹腔主干接近的横断面。</li>
		<li>将6-0 丝结扎术远端的肾上腺血管横断面放置在6.7.2。用手术剪刀将 IHVC 在左肾静脉的水平。</li>
		<li>将部分肝脏转移到后桌盘 (30 毫米培养皿), 用生理盐水填充4摄氏度。</li>
	</ol></li>
</ol>7. 后表操作 (接枝准备)
<ol><li>安装血管袖口 (图 5)<ol>
<li>用12口径的静脉注射导管用微钳将 PV 拉到袖口的腔内。用容器钳暂时固定袖口手柄和 PV。埃弗特将光伏容器壁覆盖到袖口的外表面。用圆周6-0 丝缝合固定容器壁和袖口位置。 注意: 不要拧下容器。袖口手柄应保持在12点的方向。</li>
		<li>按照7.1.1 中的说明安装 IHVC 袖口。</li>
	</ol>
</li>
	<li>将两条停留缝线预置到 SHVC 上<ol>
<li>在3点和9点方向分别使用两7-0 聚丙烯缝合线从外到内穿透 SHVC 的容器壁。 注意: 不要打结。</li>
	</ol>
</li>
	<li>保存嫁接<ol>
<li>将部分肝移植在生理盐水中浸泡4摄氏度, 以保存后表操作。</li>
	</ol>
</li>
</ol>8. 收件人操作
<ol><li>重复在 6.1-6.2 和6.7 的程序, 除了右肾切除, 在接受者暴露的手术现场, 横断面韧带和修剪血管。</li>
	<li>IHVC、胆管、肝动脉和门静脉在接受者中的制备<ol>
<li>首先, 将 IHVC 的部分调动到右肾静脉。第二, 用6-0 丝缝线结扎右肾上腺血管。第三, 利用微钳从第一肝门动员 CBD。 注意: 不要在 CBD 周围修剪脂肪组织。</li>
		<li>用6-0 丝缝线缝合双结扎术后, 在肝门的第一个分岔处横断面。然后, 用6-0 丝缝合法在双结扎术后用微剪刀对临时医管局进行横断面。将从肝门到幽门静脉的 PV 部分动员起来。</li>
	</ol></li>
	<li>调动肝脏的后方<ol>
<li>用两根棉花棒将接受者肝脏退到左侧。然后, 从 SHVC 到右肾上腺静脉切除肝脏后方的韧带, 如前一篇可视化文章18所述。在此之后, 通过 SHVC 的后空间引入橡皮筋。</li>
	</ol></li>
	<li>在无肝期阶段准备仪器和材料<ol>
<li>重新检查无肝期阶段所需的所有仪器和材料的可用性, 例如用弯曲针、微剪刀、微针支架、微钳、两个微止血钳、两7-0 聚丙烯缝合线和6-0 的5毫升注射器。真丝缝合材料。 注: 搜索材料或仪器可能会延长无肝期阶段。</li>
	</ol></li>
	<li>在接受者中切除肝脏<ol>
<li>用血管钳和直接在幽门静脉上方的 PV 交叉钳住肾脏静脉上方的 IHVC。从这个时刻记录无肝期阶段的持续时间, 直到 PV 上的钳位被释放。</li>
		<li>立即将异氟醚的浓度降低到0.5%。</li>
		<li>用预设的橡皮筋拉 SHVC 和隔膜。用斗牛犬钳夹住隔膜的部分和胸腔静脉, 保持0.5 厘米到 SHVC 的距离。 注意: 不要夹住老鼠的肺。</li>
		<li>使用微剪刀, 沿肝实质的上边缘仔细切开 SHVC 的血管壁, 尽可能长时间保持血管壁。</li>
		<li>用微剪刀对肝门第一分岔处的 PV 进行样带。然后, 对右下叶实质的下腔静脉进行横断面, 而不是在血管壁上, 将周向肝实质留在腔静脉周围。保持在右下叶的切割边缘在4毫米的距离 IHVC。transecting 血管后, 将整个肝脏从腹腔移出。</li>
	</ol></li>
	<li>重建 SHVC<ol>
<li>将部分肝移植到腹腔原位。使用两个预设的停留缝线刺穿 SHVC 的容器壁, 从内到外沿3点和9点方向。把两个留缝线拉到右边, 然后在他们被绑成一个结与自己的两端。</li>
		<li>刺穿 SHVC 血管壁从外到内接近结在3点的方向, 引入针的权利留缝线到血管腔内。</li>
		<li>吻合从3点方向到9点方向, 在血管腔内的一处连续缝合后的 SHVC 后壁。</li>
		<li>在9点方向的拐角处, 刺穿血管壁, 从内到外靠近结, 将针从血管腔中引入。</li>
		<li>使用此缝合吻合前壁使用一个正在进行的缝合线外的血管腔从9点方向到3的 o ' 时钟方向。最后, 将缝合线与它自己的末端绑3次 (图 6)。 注: 在重建的 SHVC 腔内注入3-5 毫升生理盐水, 以在最后2-3 针前强制发出气泡。SHVC 吻合的更详细的描述可以在文章中找到 Delriviere et 等。19。</li>
	</ol></li>
	<li>重建光伏发电<ol>
<li>将两个7-0 聚丙烯停留缝合在接收端的 PV 结束后, 说明在7.2。在相反的方向拉两个停留缝合, 以延长容器壁。使用两个微蚊子止血钳固定两个停留缝线。</li>
		<li>将大约1毫升的盐水注入到接收方的 PV 的血管腔内, 以强制发出气泡。快速将门静脉袖套插入到接收器中的 PV 的血管腔内, 用微钳打开。用圆周6-0 丝缝线固定袖口吻合。交叉钳。</li>
	</ol></li>
	<li>肝移植再灌注<ol>
<li>按顺序松开 SHVC 和 PV 上的夹具, reperfuse 肝移植。记录无肝期阶段的时间。检查吻合口出血。根据大鼠对疼痛刺激的反应, 将异氟醚的浓度提高到1-2%。</li>
	</ol></li>
	<li>重建 IHVC<ol>
<li>将1毫升生理盐水注入接收方 IHVC 的血管腔内, 以迫使气泡。将袖套插入 IHVC 的圆柱形血管腔内, 由圆周肝实质支撑 (图 7)。在 IHVC 的后空间引入6-0 丝缝线。用6-0 丝缝线快速固定袖口吻合。</li>
		<li>释放 IHVC 中的两个容器夹, 重新启动灌注。用微剪刀将周围的肝实质仔细修剪在圆周丝缝线上。</li>
	</ol></li>
	<li>肝动脉重建术<ol>
<li>在3点和9点方向分别使用两个11-0 聚丙烯缝合线从外到内穿透在肝移植中的茶的血管壁。咬合使用两个弯曲的微 serrefines, 在接受者中 GDA 的血流量。</li>
		<li>用微剪刀将接受者在其根部的临时医用小断面暴露在血管腔内。纵向切开部分血管壁, 以适应肝移植中的茶径, 扩大接受方临时医管局的血管腔。</li>
		<li>采用 SHVC 吻合术 (8.6. 2-8. 6.7) 所述的运行缝合技术, 将肝移植到扩大的吻合的端到端方式。 注: 详细插图可以在文章中找到黄 et al14。</li>
	</ol></li>
	<li>重建 CBD<ol>
<li>将肠道移回腹腔, 以减少胆管两端之间的距离。在接受者的胆管前壁进行横切口。</li>
		<li>将胆道支架拉入部分肝移植物中, 通过横切口将其插入到接受者胆管的腔内。用6-0 周丝缝线固定支架。</li>
		<li>将两个6-0 周围丝缝线缝合在胆管之间, 以减少吻合的张力。</li>
	</ol></li>
	<li>关闭腹部<ol>
<li>在连续缝合模式下用3-0 丝缝线缝合两层腹部。</li>
	</ol></li>
</ol>9. 术后治疗 (镇痛和抗生素)
<ol><li>治疗所有接受者0.1 毫克/千克丁丙诺啡皮下。治疗头孢呋辛 (16 毫克/千克) 除了止痛药。向老鼠提供自来水和标准实验动物周向ad 随意。</li>
</ol>

<ol>
	<li>Yan, J. Q., Becker, T., Peng, C. H., Li, H. W., Klempnauer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Split+liver+transplantation:+a+reliable+approach+to+expand+donor+pool.">Split liver transplantation: a reliable approach to expand donor pool.</a> Hepatobiliary Pancreat Dis Int. 4, 339-344 (2005).</li><li>Xia, R., Emond, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+partial+liver+transplantation+in+the+rat:+a+model+of+70%+hepatectomy+and+reduced+size+liver+transplantation.">Orthotopic partial liver transplantation in the rat: a model of 70% hepatectomy and reduced size liver transplantation.</a> Transplantation. 56, 1041-1043 (1993).</li><li>Omura, T., Ascher, N. L., Emond, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fifty-percent+partial+liver+transplantation+in+the+rat.">Fifty-percent partial liver transplantation in the rat.</a> Transplantation. 62, 292-293 (1996).</li><li>Foss, A., 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=Function+of+reduced-size+liver+transplant+in+the+rat.">Function of reduced-size liver transplant in the rat.</a> Res Exp Med (Berl). 197, 91-99 (1997).</li><li>Nagai, K., Yagi, S., Uemoto, S., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+procedures+for+a+rat+model+of+partial+orthotopic+liver+transplantation+with+hepatic+arterial+reconstruction.">Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction.</a> J Vis Exp. , e4376 (2013).</li><li>Sun, J., Qin, G., Wu, L., Wang, C., Sheil, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antigenic+load+and+peripheral+chimeric+levels+in+entire+and+partial+liver+allograft+recipients.">Antigenic load and peripheral chimeric levels in entire and partial liver allograft recipients.</a> Microsurgery. 21, 183-187 (2001).</li><li>Ninomiya, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deceleration+of+regenerative+response+improves+the+outcome+of+rat+with+massive+hepatectomy.">Deceleration of regenerative response improves the outcome of rat with massive hepatectomy.</a> Am J Transplant. 10, 1580-1587 (2010).</li><li>Yamanaka, K., 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=Effect+of+olprinone+on+liver+microstructure+in+rat+partial+liver+transplantation.">Effect of olprinone on liver microstructure in rat partial liver transplantation.</a> J Surg Res. 183, 391-396 (2013).</li><li>Wang, W., 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=Mesenchymal+stem+cells+promote+liver+regeneration+and+prolong+survival+in+small-for-size+liver+grafts:+involvement+of+C-Jun+N-terminal+kinase,+cyclin+D1,+and+NF-kappaB.">Mesenchymal stem cells promote liver regeneration and prolong survival in small-for-size liver grafts: involvement of C-Jun N-terminal kinase, cyclin D1, and NF-kappaB.</a> PLoS One. 9, e112532 (2014).</li><li>Madrahimov, N., Dirsch, O., Broelsch, C., Dahmen, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marginal+hepatectomy+in+the+rat:+from+anatomy+to+surgery.">Marginal hepatectomy in the rat: from anatomy to surgery.</a> Ann. Surg. 244, 89-98 (2006).</li><li>Martins, P. N., Theruvath, T. P., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+partial+hepatectomies.">Rodent models of partial hepatectomies.</a> Liver Int. 28, 3-11 (2008).</li><li>Kamada, N., Sumimoto, R., Kaneda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+value+of+hepatic+artery+reconstruction+as+a+technique+in+rat+liver+transplantation.">The value of hepatic artery reconstruction as a technique in rat liver transplantation.</a> Surgery. 111, 195-200 (1992).</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, 327-332 (2003).</li><li>Huang, 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=A+novel+end-to-side+anastomosis+technique+for+hepatic+rearterialization+in+rat+orthotopic+liver+transplantation+to+accommodate+size+mismatches+between+vessels.">A novel end-to-side anastomosis technique for hepatic rearterialization in rat orthotopic liver transplantation to accommodate size mismatches between vessels.</a> Eur Surg Res. 47, 53-62 (2011).</li><li>Bernal, 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=Guidelines+for+rodent+survival+surgery.">Guidelines for rodent survival surgery.</a> J.Invest Surg. 22, 445-451 (2009).</li><li>Johnson, A. J., Stulting, R. D., Macsai, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ophthalmic Microsurgical Suturing Techniques. , 21-28 (2007).</li><li>Delriviere, L., 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=Detailed+modified+technique+for+safer+harvesting+and+preparation+of+liver+graft+in+the+rat.">Detailed modified technique for safer harvesting and preparation of liver graft in the rat.</a> Microsurgery. 17, 690-696 (1996).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <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+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp. , (2012).</li><li>Delriviere, L., 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=Technical+details+for+safer+venous+and+biliary+anastomoses+for+liver+transplantation+in+the+rat.">Technical details for safer venous and biliary anastomoses for liver transplantation in the rat.</a> Microsurgery. 18, 12-18 (1998).</li><li>Jin, 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=Preliminary+experience+of+a+PDCA-cycle+and+quality+management+based+training+curriculum+for+rat+liver+transplantation.">Preliminary experience of a PDCA-cycle and quality management based training curriculum for rat liver transplantation.</a> J Surg Res. 176, 409-422 (2012).</li><li>Selzner, N., Selzner, M., Tian, Y., Kadry, Z., 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=Cold+ischemia+decreases+liver+regeneration+after+partial+liver+transplantation+in+the+rat:+A+TNF-alpha/IL-6-dependent+mechanism.">Cold ischemia decreases liver regeneration after partial liver transplantation in the rat: A TNF-alpha/IL-6-dependent mechanism.</a> Hepatology. 36, 812-818 (2002).</li><li>Tanaka, H., Hashizume, K., Enosawa, S., Suzuki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+a+20%+partial+liver+graft+in+rats:+a+technical+innovation.">Successful transplantation of a 20% partial liver graft in rats: a technical innovation.</a> J Surg Res. 110, 409-412 (2003).</li><li>Bockhorn, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Erythropoietin+treatment+improves+liver+regeneration+and+survival+in+rat+models+of+extended+liver+resection+and+living+donor+liver+transplantation.">Erythropoietin treatment improves liver regeneration and survival in rat models of extended liver resection and living donor liver transplantation.</a> Transplantation. 86, 1578-1585 (2008).</li><li>Martins, P. N., Neuhaus, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+anatomy+of+the+liver,+hepatic+vasculature+and+bile+ducts+in+the+rat.">Surgical anatomy of the liver, hepatic vasculature and bile ducts in the rat.</a> Liver Int. 27, 384-392 (2007).</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, 47-50 (1979).</li><li>Kashfi, A., 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+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplant.Proc. 37, 185-188 (2005).</li><li>Wain, R. A. J., Hammond, D., McPhillips, M., Whitty, J. P. M., Ahmed, W., Ahmed, W., Jackson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Surgical Tools and Medical Devices. , 545-562 (2016).</li><li>Huang, 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=Hepatic+arterial+perfusion+is+essential+for+the+spontaneous+recovery+from+focal+hepatic+venous+outflow+obstruction+in+rats.">Hepatic arterial perfusion is essential for the spontaneous recovery from focal hepatic venous outflow obstruction in rats.</a> Am J Transplant. 11, 2342-2352 (2011).</li><li>Czigany, Z., 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=Improving+Research+Practice+in+Rat+Orthotopic+and+Partial+Orthotopic+Liver+Transplantation:+A+Review,+Recommendation,+and+Publication+Guide.">Improving Research Practice in Rat Orthotopic and Partial Orthotopic Liver Transplantation: A Review, Recommendation, and Publication Guide.</a> Eur Surg Res. 55, 119-138 (2015).</li></ol>

所有作者都没有相互冲突的利益来披露。

大鼠 PLTx 是一种复杂的显微外科手术, 其培训计划成本高且持续时间为20。大鼠 PLTx 协议的复杂性使得研究人员无法使用这种动物模型。与全尺寸大鼠 LTx 相比, 大鼠 PLTx 的 microsurgeon 不仅与移植过程的挑战有关, 而且与小动物肝脏切除的挑战有关。因此, 详细描述整个过程的可视化显微外科协议是建立这一复杂模型的必要条件。Nagai et所报告的先前可视化的文章提出了一项50%...

目前, 捐赠肝脏的数量和等待捐献肝脏的病人数量之间存在很大的差异。肝脏器官的短缺是一个全球性的问题。为了扩大捐献者的水池, 分裂肝移植 (LTx) 和活体捐献者 LTx 被开发用于使用部分肝脏作为移植物1。
为进一步研究部分肝移植 (PLTx) 的机制, 建立了相关动物模型2,3,4,5。在大鼠 PLTx 中, 肝叶切除体内和前体, 以模拟活体捐献者 LTx 的条件, 并分别在人类体内分裂 LTx。在可视化实验杂志上发表的一篇论文提出了一个详细的协议, 涉及50% 只老鼠 PLTx 使用前体肝切除5。然而, 在可视化文献中还没有报道一只老鼠 PLTx 与体内肝切除术有关。
除了体内和前体hepatectomies 之间的差异外, 执行肝切除术本身也在确定 PLTx 的结果方面起着重要作用. 目前, 在许多外科研究中使用大鼠 PTLx模型, 肝叶切除后, 放置一个简单的结扎在肝瓣的椎弓根2,3,6,7,8,9。然而, 在切除前放置一个简单的结扎不适合所有肝叶, 因为不同的肝叶有不同的形状和大小。在正中裂片的基础上进行简单结扎, 有很高的风险导致腔静脉收缩, 这可能最终影响部分肝移植的流出10,11。因此, 在大鼠 PTLx 的研究中, 需要以大鼠肝脏解剖知识为基础的更新肝切除技术。
在本研究中所述的协议中, 将最新的以血管为导向的70% 肝切除术纳入大鼠 PLTx 的程序中。在肝实质切除前分别对左外侧叶 (微光) 和正中叶 (ML) 的门静脉和肝动脉进行解剖和分切。然后用穿刺缝线结扎了微光和 ML 的肝静脉。通过使用单独的结扎和多重穿刺缝合, 而不是简单的结扎, 毫升的残余树桩能够扩散到腔静脉。因此, 避免了手术结扎引起的 infrahepatic 腔静脉的收缩。另外, 在切除肝实质之前, 单独结扎对微光和 ML 血供的阻断减少了残肝残端的出血率, 从而最大限度地减少了血液损失对实验的影响11。
对于 microsurgeons, 由于该血管直径极小, 重建肝移植肝动脉 (PHA) 是一项重大的挑战。虽然在 LTx 中重新动脉是否真的是必要的问题仍在辩论中12,13,14, 已经提出了许多重建肝动脉的显微外科技术14.本文介绍了一种新的肝移植动脉技术, 它以端到端的方式将普通肝动脉 (CHA) 与扩大后的医用血管吻合。使用这种端对端技术, 两个肝动脉连接到一个更大的直径吻合。吻合口直径越大, 缝合越容易, 吻合通畅的改善越大。

<table><tbody><tr><td>Surgical microscope</td><td>M&amp;ouml;ller-Wedel, Germany</td><td>654359</td><td></td></tr><tr><td>Light source</td><td>OSRAM (China) Lighting Co., Ltd</td><td>64627</td><td></td></tr><tr><td>Isoflurane Vaporizer</td><td>MIDMARK Corporation</td><td>VIP3000</td><td></td></tr><tr><td>Isoflurane</td><td>Baxter Healthcare Corporation</td><td>40032609</td><td>For Inhalation anesthesia</td></tr><tr><td>Normal Saline</td><td>Zhejiang Tianrui Pharmaceutical Co., Ltd.</td><td>716092103</td><td></td></tr><tr><td>Povidone Iodine Solution</td><td>HANGZHOU MINSHENG PHARMACEUTICAL</td><td>H33021567</td><td>For disinfection</td></tr><tr><td>Hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J31020</td><td></td></tr><tr><td>Surgical scissors</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>JC2116</td><td></td></tr><tr><td>Needle-holder</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J32030</td><td></td></tr><tr><td>Dressing Forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>J42020</td><td></td></tr><tr><td>Mosquito hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>W40340</td><td>For liver resection; For retracting the stay sutures.&amp;nbsp;</td></tr><tr><td>Micro-scissors</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA1040</td><td></td></tr><tr><td>Micro needle-holder</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA2060</td><td></td></tr><tr><td>Curved micro typing forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3061</td><td></td></tr><tr><td>straight micro typing forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3060</td><td></td></tr><tr><td>Micro hemostatic forceps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>WA3020</td><td></td></tr><tr><td>Bulldog clamp</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>XEC350</td><td></td></tr><tr><td>Vessel clamps</td><td>Shanghai Medical Instruments(Group) Ltd</td><td>W40160</td><td></td></tr><tr><td>Micro Serrefine-curved</td><td>&amp;nbsp;Fine Science Tools, Inc</td><td>18055-05</td><td></td></tr><tr><td>Dacryosyrinx</td><td>Shi Zhuo medical instruments Co., Ltd.</td><td>5# curve</td><td>Use as the curved needle for forcing out the air bubbles in the vascular lumen</td></tr><tr><td>Sterilized gauze swabs</td><td>Fuqing Health &amp;amp; Integral Medical</td><td>20230R</td><td></td></tr><tr><td>5mL syringe</td><td>Zhejiang Yusheng Medical Instrument Co.,Ltd</td><td>811932416</td><td></td></tr><tr><td>18G needles</td><td>Shanghai Kindly Enterprise Development Group Co., Ltd</td><td>01-014</td><td>For fixing the upper abdomen wall</td></tr><tr><td>Intima II 22G intraveous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>383207y</td><td>For making biliary stent; For infusing perfusate</td></tr><tr><td>7-0 polypropylene sutures</td><td>Ningbo Medical Needle Co., Ltd</td><td>151026</td><td>For SHVC anastomsis</td></tr><tr><td>6-0 silk suture</td><td>Shanghai Pudong Jinhuan Medical Products Co., Ltd</td><td>2650182</td><td>For ligation</td></tr><tr><td>Culture dish (100mm)</td><td>Corning Incorporated</td><td>430165</td><td>used in back table for storage ice. The back table dish was placed on it.&amp;nbsp;</td></tr><tr><td>Culture dish (30 mm)</td><td>Corning Incorporated</td><td>3296</td><td>As a vessel for organ preservation</td></tr><tr><td>Angiocath 12G intravenous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>382277</td><td>For making a IHVC cuff</td></tr><tr><td>Angiocath 14G intravenous catheter</td><td>Becton Dickinson Medical Devices (Shanghai) Co Ltd.</td><td>382269</td><td>For making a PV cuff</td></tr><tr><td>Buprenorphine (hydrochloride)</td><td>Tianjin Institute of Pharmaceutical Research Pharmaceutical Co., Ltd</td><td>H12020275</td><td>For analgesic</td></tr><tr><td>Cefuroxime sodium for injection</td><td>Esseti FarmaceuticiS.r.l</td><td>H20160013</td><td>As an antibiotics</td></tr></tbody></table>

re-arterialized rat partial liver transplantation with an in vivo vessel-oriented 70% hepatectomy

在临床上发展了肝移植和活体肝供肝移植, 以更有效地利用肝脏器官。为更好地了解这些手术过程的机制, 建立了大鼠部分肝移植 (PLTx) 模型进行相关的外科研究。由于大鼠 PLTx 模型的复杂性, 需要一个具有详细描述的协议。此前发表的一篇文章报告了一项协议, 其中前体肝切除术用于实现50% 大鼠 PLTx。与此协议相比, 我们引入了一个 PLTx体内70% 肝切除的重新静脉。以血管为导向的肝切除术被纳入大鼠 PLTx, 以改进显微外科手术。肝实质切除前, 左外侧叶和正中叶的门静脉和肝动脉分别进行解剖和结扎, 从而降低残肝残端出血的几率。此外, 还介绍了肝动脉与肝动脉扩张的端到端血管吻合术, 以重新 arterialize 肝脏动脉。采用端到端血管吻合技术, 扩大了吻合口的直径, 降低了手缝的难度, 保持了较高的吻合通畅率。此外, infrahepatic 腔静脉的袖套吻合略有改变。在袖状吻合术期间, 在接受者的腔静脉周围有一节环性肝实质, 以维持血管腔的三维形状。这部分肝实质在完成吻合术后被切除。经此修改, 省略了放置缝合线的步骤, 从而进一步缩短了袖套吻合时间。采用本协议对大鼠 PLTx, 在显微外科手术后实现了低肝酶水平、完整的肝小叶结构和高存活率。

共完成31例自体静脉大鼠 PLTx 的应用本协议。所有收件人都存活到观察时间结束为止。患者的体重在术后一天 (POD) 4 开始恢复。在 POD 6 (图 8) 之后, 体重的斜率接近正常的 Lewis 鼠, 这间接地暗示了接收方的恢复。组织学上, 接受者观察到胆管的轻微增生, 接受者的肝小叶结构完好。没有发现坏死或明显的正弦扩张 (图 9)。血清丙氨...

Watch this Scientific Journal Video about Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy at JoVE.com

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

本文提出了一种静脉大鼠部分肝移植的协议。具体地说, 70% 例肝切除术是通过使用一种最新的血管性肝切除技术在体内进行的。肝动脉以端对侧的方式重建。对袖套技术进行了改进, 缩短了 infrahepatic 腔静脉的吻合时间。

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. ...

re-arterialized-rat-partial-liver-transplantation-with-an-vivo-vessel

Research

JoVE Journal

Medicine

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

11.5K 次观看.  The First Affiliated Hospital of Wenzhou Medical University. 本文提出了一种静脉大鼠部分肝移植的协议。具体地说, 70% 例肝切除术是通过使用一种最新的血管性肝切除技术在体内进行的。肝动脉以端对侧的方式重建。对袖套技术进行了改进, 缩短了 infrahepatic 腔静脉的吻合时间。

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

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 ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

大鼠体内部分肝工程的外科技术基础

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

生物工程

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the silicone compound, via a 26 G catheter, into the portal or hepatic vein. After silicone injection, organs were explanted and prepared for ex-vivo micro-CT (&#181;CT) scanning. The silicone injection procedure is technically challenging. Achieving a successful outcome requires extensive microsurgical experience from the surgeon. One of the challenges of this procedure involves determining the adequate perfusion rate for the silicone compound. The perfusion rate for the silicone compound needs to be defined based on the hemodynamic of the vascular system of interest. Inappropriate perfusion rate can lead to an incomplete perfusion, artificial dilation and rupturing of vascular trees.
The 3D reconstruction of the vascular system was based on CT scans and was achieved using preclinical software such as HepaVision. The quality of the reconstructed vascular tree was directly related to the quality of silicone perfusion. Subsequently computed vascular parameters indicative of vascular growth, such as total vascular volume, were calculated based on the vascular reconstructions. Contrasting the vascular tree with silicone allowed for subsequent histological work-up of the specimen after &#181;CT scanning. The specimen can be subjected to serial sectioning, histological analysis and whole slide scanning, and thereafter to 3D reconstruction of the vascular trees based on histological images. This is the prerequisite for the detection of molecular events and their distribution with respect to the vascular tree. This modified silicone injection procedure can also be used to visualize and reconstruct the vascular systems of other organs. This technique has the potential to be extensively applied to studies concerning vascular anatomy and growth in various animal and disease models.

Tools used for visualizing vascular regeneration require methods for contrasting the vascular trees. This film demonstrated a delicate injection technique used to achieve optimal contrasting of the vascular trees and illustrate the potential benefits resulting from a detailed analysis of the resulting specimen using &#181;CT and histological serial sections.

之后，正常小鼠70％部分肝切除血管和实质再生的可视化

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...

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 ...

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, ...

小鼠肝移植中有效血管吻合的策略

Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. However, the technical challenges pose a barrier to utilizing this valuable experimental model and passing on these skills to the next generation. The most challenging aspect of this procedure is vascular reconstruction, including the portal vein (PV), infrahepatic inferior vena cava (IHIVC), and suprahepatic inferior vena cava. The use of plastic cuffs, rather than sutures, allows for smoother PV and IHIVC reconstruction. Vessels are reconstructed by attaching a cuff made from an intravenous catheter to the tip of the graft vessel and interposing the cuff into the recipient vessel. The two most crucial aspects are properly visualizing the inner lumen of the vessel and avoiding the use of excessive force. Our aim is to provide a technical overview of vascular reconstructions using the cuff technique in recipient surgery. These technical tips for the cuff technique are expected to help microsurgeons facilitate vascular reconstruction and advance their research.

作者聚焦：探索脂肪肝移植中的冷缺血和温再灌注损伤

This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.

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. ...

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

摘要

探索更多视频

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

大鼠体内部分肝工程的外科技术基础

之后，正常小鼠70％部分肝切除血管和实质再生的可视化

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

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

袖带技术在小鼠原位肝移植中的血管重建

袖带技术在小鼠原位肝移植中的血管重建