这项研究经费由“德意志研究联合会”（DFG）。 K. Schleimer是由国家科技部的莉泽·迈特纳北威州奖学金和研究支持。我们要感谢夫人玛丽 - 琼Blümich为她优秀的编辑协助。玛丽亚Kokozidou博士从费森尤斯2011_A61资金。

1，实验动物<ol><li>获得了将要执行的，依照动物保护FELASA指引所有实验当地伦理委员会的批准（K 40，17/99）。 </li><li>用雄性Lewis大鼠可从任何商业育种（ 如查尔斯河Wiga有限公司，的Sulzfeld，德国）购买。这里，224只动物（112供体，112的收件人）被用来执行112移植。处死术后6周收件人。在操作时，将动物应权衡250-350克。 </li><li>请在空调房的动物免费获得食物和水。 </li></ol> 2，麻醉和手术准备<ol><li>将大鼠在一个有机玻璃盒连接到麻醉机用的1升/分钟的氧气流率，和一个异氟烷蒸发器，用5％的麻醉气体，以诱导麻醉。麻醉维持在1.5％异氟醚。 </li><li>将大鼠在保温板仰卧位。 </li><li>用脱毛霜去除腹部的皮毛。 </li><li>准备一个无菌手术领域如下：使用消毒剂来清洁台面。对于切口部位用氯己定的杀菌第一，醇秒，最后，擦洗溶液（ 例如，氯己定和70％的异丙醇）。穿一次性手术衣和手术手套。 </li><li>使用无菌显微仪器。 </li><li>使用12-20倍放大倍率的显微镜操作。 </li></ol> 3，手术过程<ol><li>供体手术 准备 <ol><li>执行中位数剖腹和切除左外叶和所描述的希金斯移植物的平均叶14 </li><li>然后准备肝十二指肠韧带。第一结扎胆管，以诱导充血的小胆管和扩张。然后结扎和横切gastroduodeNAL静脉和动脉。此后，隔离腹腔干和肝总动脉和横切胃左动脉和脾动脉。切开拥挤的胆管，并引入20-G支架进去。支架的定位必须确保在整个肝门部胆管系统的引流。样远端胆管的扩张。 </li><li>隔离腔静脉的肝下段正上方右侧肾静​​脉。 灌注 </li><li>上述腹腔干主动脉结扎后，用镊子将门静脉和切割它。引入灌注导管（14 G）的进入门静脉，并用30ml的组氨酸 - 色氨酸 - 酮戊二酸（HTK） - 解决方案（12厘米水柱，4℃）进行灌注。 </li><li>切开隔膜和肝上腔静脉，以允许灌注液流出。 </li><li>切除腹腔干与主动脉修补和组氨酸，色氨酸-酮戊二酸溶液冲洗。 外植 </li><li>样的肝下腔静脉正上方右侧肾静​​脉。 </li><li>然后，将支架与门静脉直径0.3毫米，用绷带固定。在下面的操作中，门静脉经由右肾动脉动脉化。远端样的门静脉支架。 </li><li>肝上下腔静脉结扎后，有隔膜的袖口横切在一起。 </li><li>外植体移植物，并在4摄氏度，它的组氨酸，色氨酸-酮戊二酸（HTK） - 解决方案存储在一个塑料袋上的冰。广场一角缝线（8-0 Ethilon）在肝下腔静脉。 </li></ol></li><li>受体手术<ol><li>执行中位数剖腹探查，并准备肝下腔静脉。然后分离出右肾动脉和输尿管，结扎输尿管和横切它。用镊子将右肾动脉的起源，然后结扎右肾静脉。那么两者的横断后重新NAL船，执行切除术。随后，缩短了右肾动脉，用肝素盐水（40学时肝素/毫升生理盐水溶液）冲洗。 </li><li>近端样右肾静脉至肝下下腔静脉的近端和远端的夹紧后结扎在肝下下腔静脉创建一个椭圆形的腔。冲洗腔静脉用肝素生理盐水溶液。植入收件人的腹部右侧肾区移植。 </li><li>吻合的捐赠者的肝下下腔静脉端 - 侧到recipient`s腔静脉用8-0 Ethilon连续缝合。 </li><li>用肝素/盐水溶液冲洗后，介绍在门静脉支架插入右侧肾动脉（ 图3）。连接门静脉和肾动脉与2针（8-0 Ethilon）。修复肾动脉支架连接用绷带（编织丝黑7-0）。 </li><li>要允许再灌注，先取下动脉夹，那么这两个静脉夹（ 图4）。 </li><li>准备下腹主动脉，然后将近端和远端微夹钳和切开右两个夹子之间的主动脉。进行主动脉和捐赠者的腹腔干之间的端 - 侧吻合术，采用主动脉修补，以10-0 Ethilon连续缝合。 </li><li>插入胆管进入十二指肠：在十二指肠壁涂抹pursestring缝合，做一个小切口，在十二指肠壁就在pursestring的中间。引进支架胆管进入十二指肠，并拧紧pursestring缝合。最后，保持十二指肠和总胆管紧紧连同两针（Ethilon 8-0）（ 图5）。 </li><li>然后进行次全切除原生肝，首先由Emond 等人 。模仿急性肝功能衰竭。7 </li><li>修复移植的肝上下腔静脉，膈肌的袖口在一起，重新的侧腹壁cipient用4-0薇乔缝合。冲洗用生理盐水溶液（ 图6）的腹部。 </li><li>对于肌层用4-0薇乔连续缝合，并为皮肤3-0薇乔连续缝合。 </li></ol></li></ol> 4，恢复和术后管理<ol><li>让动物从麻醉下红外加热灯进行恢复。 </li><li>注射丁丙诺啡在2天剂量为0.03-0.05毫克/千克体重SC每8-12小时。 </li><li>审议有关的一般情况和疾病或感染的迹象的动物。 </li></ol>初始本土肝脏重量的5计算计算初始天然肝重量以下面的方式：从完整的肝脏重量中减去切除肝叶的重量（体重（此数值为±0.49％的标准偏差为2.66％是在以前的实验中有n个判定= 80动物一脉相承和中GE））15

<ol>
	<li>Boudjema, 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=Auxiliary+liver+transplantation+for+fulminant+and+subfulminant+hepatic+failure.">Auxiliary liver transplantation for fulminant and subfulminant hepatic failure.</a> Transplantation. 59, 218-223 (1995).</li><li>Boudjema, K., Jaeck, D., Sim&#233;oni, U., Bientz, J., Chenard, M. P., Brunot, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporary+auxiliary+liver+transplantation+for+subacute+liver+failure+in+a+child.">Temporary auxiliary liver transplantation for subacute liver failure in a child.</a> The Lancet. 342, 778-779 (1993).</li><li>Chenard-Neu, M. P., 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=Auxiliary+liver+transplantation:+Regeneration+of+the+native+liver+and+outcome+in+30+patients+with+fulminant+hepatic+failure+-+a+multicenter+European+study.">Auxiliary liver transplantation: Regeneration of the native liver and outcome in 30 patients with fulminant hepatic failure - a multicenter European study.</a> Hepatology. 23, 1119-1127 (1996).</li><li>Gubernatis, G., Pichlmayr, R., Kemnitz, J., Gratz, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver&#8211;transplantation+(APOLT)+for+fulminant+hepatic+failure:+first+successful+case+report.">Auxiliary partial orthotopic liver&#8211;transplantation (APOLT) for fulminant hepatic failure: first successful case report.</a> World J Surg. 15, 660-666 (1991).</li><li>Erhard, J., Lange, R., Giebler, R., Rauen, U., de Groot, H., Eigler, F. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+grafting:+a+new+concept+in+liver+transplantation.">Auxiliary liver grafting: a new concept in liver transplantation.</a> The Lancet. 342, 758 (1993).</li><li>Terpstra, O. T., Reuvers, C. B., Schalm, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+heterotopic+liver+transplantation.">Auxiliary heterotopic liver transplantation.</a> Transplantation. 45, 1003-1007 (1988).</li><li>Schleimer, K., Lange, R., Rauen, U., Erhard, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+in+acute+liver+failure+in+the+rat+-+an+illustrated+description+of+a+new+surgical+approach.">Auxiliary liver transplantation in acute liver failure in the rat - an illustrated description of a new surgical approach.</a> Langenbecks Arch Surg. 384 (2), 204-208 (1999).</li><li>Schleimer, 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=Physiologic+microcirculation+of+the+heterotopically+transplanted+rat+liver+with+portal+vein+arterialization+depending+on+optimal+stent+diameter.">Physiologic microcirculation of the heterotopically transplanted rat liver with portal vein arterialization depending on optimal stent diameter.</a> Med Sci Monit. 28 (BR140-145), (2006).</li><li>Hong, H. Q., 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=Study+of+portal+arterialization+with+auxiliary+liver+in+rats.">Study of portal arterialization with auxiliary liver in rats.</a> Hiroshima J. Med. Sci. 40, 29-33 (1991).</li><li>Higgins, G. M., Anderson, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+pathology+of+the+liver.+I--Restoration+of+the+liver+of+the+white+rat+following+partial+surgical+removal.">Experimental pathology of the liver. I--Restoration of the liver of the white rat following partial surgical removal.</a> Arch. Path. 12, 186-202 (1931).</li><li>Schleimer, 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=Auxiliary+liver+transplantation+with+flow-regulated+portal+vein+arterialization+offers+a+successful+therapeutic+option+in+acute+hepatic+failure--investigations+in+heterotopic+auxiliary+rat+liver+transplantation.">Auxiliary liver transplantation with flow-regulated portal vein arterialization offers a successful therapeutic option in acute hepatic failure--investigations in heterotopic auxiliary rat liver transplantation.</a> Transpl Int. 19 (7), 581-588 (2006).</li><li>Lee, S., Edgington, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+transplantation+in+the+rat.">Liver transplantation in the rat.</a> Surg. 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Res. 9, 270-279 (1977).</li><li>Aguirrezabalaga, J., Arnal, F., Marini, M., Centeno, A., Fernandez-Selles, C., Rey, I., Gomez, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+liver+transplantation+with+portal+arterialization+in+the+rat:+description+of+a+new+model.">Auxiliary liver transplantation with portal arterialization in the rat: description of a new model.</a> Microsurgery. 22, 21-26 (2002).</li><li>Palmes, D., Dietl, K. H., Drews, G., Holzen, J. P., Spiegel, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Auxiliary+partial+orthotopic+liver+transplantation:+treatment+of+acute+liver+failure+in+a+new+rat+model.">Auxiliary partial orthotopic liver transplantation: treatment of acute liver failure in a new rat model.</a> Langenbecks Arch Surg. (386), 534-541 (2002).</li><li>Schleimer, 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=Improved+microcirculation+of+a+liver+graft+by+controlled+portal+vein+arterialization.">Improved microcirculation of a liver graft by controlled portal vein arterialization.</a> J Surg Res. 116 (2), 202-210 (2004).</li><li>Schleimer, 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=Portal+hyperperfusion+causes+disturbance+of+microcirculation+and+increased+rate+of+hepatocellular+apoptosis:+investigations+in+heterotopic+rat+liver+transplantation+with+portal+vein+arterialization.">Portal hyperperfusion causes disturbance of microcirculation and increased rate of hepatocellular apoptosis: investigations in heterotopic rat liver transplantation with portal vein arterialization.</a> Transplant Proc. 38 (3), 725-729 (2006).</li></ol>

所有作者宣称没有利益冲突。

第一异位辅助大鼠肝移植是由Lee 等人所描述的，在1966年16接着赫斯。移植物17中的门静脉吻合与接受者的门静脉。在此过程中，门静脉血液供应天然肝脏受到损害。 1991年，香港通过收件人的髂内动脉进行暂停与PVA的13无血流调节大规模门户网站和血窦拥塞的发生。 此外Aguirrezabalaga 等 ，进行暂停与PVA，但没有...

急性肝功能衰竭造成临床医学的一个严重问题，导致死亡率为60％-80％。8在急性肝功能衰竭，辅助肝移植是一种很有前途的替代疗法来弥补急性期。原生肝左再生和下面的再生免疫抑制药物可停药。作为结果，移植将成为萎缩。然而，手术技术辅助肝移植仍然是讨论。1-3在此背景下，特普斯特拉报空间不足的移植，门脉血盗血现象和静脉淤血和排水的问题。9-10捐赠者的门静脉吻合的端至端向接收方的门静脉。这导致在接枝和天然肝之间和在门静脉血栓形成门脉血流竞争。大多数的这些技术问题，可避免与使用APOLT-法（orthotopIC辅助段植入）。4但是这种方法需要两个原生肝移植。4在1998年广泛切除，艾哈德和他的团队开发了异位辅助肝移植（暂停）利用门静脉动脉聚乙烯醇（PVA）（ 图1 ）。肺门和本地肝脏的门静脉受到尊重与使用这种方法。 ，不遗余力，从不必要的副作用，如门静脉血栓形成和门脉血收件人盗取的现象，这可能会扰乱当地的肝脏恢复。此外，还需要对这个方法的原生肝移植无肝广泛切除，因而具有出血少，更其余全肝实质质量桥接（移植）和再生（原生肝）。这种技术表现出乐观的初步临床结果。5-6本实验工作的目标是暂停技术与门静脉动脉化发展大鼠用于研究目的（ 图2）。在初步实验中，我们并没有考虑到动脉化门静脉血流调节静脉，导致再灌注损伤和大出血的肝包膜充血后移植移植11此后在动脉化门静脉血流办法，由支架为0.3mm，内径为8毫米的长度。试点试验已经表明，在0.3毫米的支架直径得到的平均血流在动脉化门静脉即上部生理范围内（1.7±0.4毫升/分钟/克肝脏重量）12我们没有血流调节的结果与香港，谁通过收件人的髂内动脉进行暂停与PVA的13在他的实验中，没有血流调节大量的门户网站和正弦波发生拥塞的结果相关。 我们的视频的目的是与demonstrate步步这个ne.w实验外科技术在大鼠。在长期的提出的技术可以在肝脏再生和跨肝脏的竞争和门静脉动脉，门静脉高灌注及门脉血流调控研究中使用。

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		<tr height="119">
			<td height="119" style="height:119px;width:216px;">HTK solution,</td>
			<td rowspan="2" style="width:71px;">Dr. Franz K&#246;hler Chemie GmbH, Alsbach-H&#228;hnlein, Germany</td>
			<td rowspan="2" style="width:119px;"></td>
			<td rowspan="2" style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Custodiol</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Isofluorane 5 %</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Heparin</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">0.9% saline</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Buprenorphine</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Any genericon</td>
		</tr>
	</tbody>
</table>

heterotopic auxiliary rat liver transplantation with flow-regulated portal vein arterialization in acute hepatic failure

在急性肝功能衰竭的辅助肝移植是一个有趣的替代方法。其目的在于，提供一种临时支撑，直到发生故障的原生肝已再生。1-3 APOLT法，辅助segments-的原位移植可避免出现最多的技术问题。然而此方法需要两个天然肝和接枝。4在1998年广泛切除，艾哈德开发的异位辅助肝移植（HALT）利用门静脉动脉（PVA）（图1）。该技术表现出有前途的初步临床结果。5-6，我们开发出了HALT-技术与流量调节的PVA在大鼠研究流动调节PVA的上接枝形态和功能（图2）的影响。 一种肝移植缩小到原始大小的30％，是右肾外植后异位植入接受者的右侧肾区。60;移植物的红外肝腔静脉吻合与接受者的肝下腔静脉。捐赠者的门静脉动脉化进行了通过收件人的右肾动脉支架技术。动脉化门静脉的血流量的调节是通过使用一个支架上以0.3mm的内部直径来实现。移植物的腹腔干为端 - 侧吻合用收件人的主动脉和胆管被植入到十二指肠。执行了次全切除原生肝诱发急性肝功能衰竭。7 以这种方式112移植被执行。围手术期的存活率为90％和6周的存活率为80％。术后6周，天然肝再生，表示重量的增加，从2.3±0.8克至9.8±1克此时，接枝的重量从3.3±0.8克下降至2.3±0.8克</p&gt; 我们能够在接枝形态和功能的角度，以获得希望的长期结果。暂停与流量调节PVA的可靠桥梁急性肝功能衰竭，直至原生的肝脏再生。

移植的平均时间约为150分钟。冷缺血时间为64±7分钟时，热缺血时间为小于25分钟。 围手术期的存活率为90％和6周的存活率为80％。在术后第一周的20％的损失是由于因支架错位和胆漏（4只）腹膜炎，由于在接下来的5周肺炎，腹腔内脓肿及肠梗阻（7只）。幸存的动物们在良好的一般状况。 肝脏合成的参数（连结值：110±7.8％，AT III：104±6％）操作六周后?...

Watch this Scientific Journal Video about Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure at JoVE.com

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

辅助性肝移植提供了急性肝功能衰竭的临时支撑，直到失败的肝脏再生。异位辅助肝移植（HALT）与门静脉动脉聚乙烯醇（PVA）呈现足够的肝功能。我们开发了一种类似的方法在大鼠中，以检查对移植物的形态和功能的门静脉动脉化的影响。

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 ...

heterotopic-auxiliary-rat-liver-transplantation-with-flow-regulated

Research

JoVE Journal

Medicine

12.6K 次观看.  University Hospital RWTH Aachen. 辅助性肝移植提供了急性肝功能衰竭的临时支撑，直到失败的肝脏再生。异位辅助肝移植（HALT）与门静脉动脉聚乙烯醇（PVA）呈现足够的肝功能。我们开发了一种类似的方法在大鼠中，以检查对移植物的形态和功能的门静脉动脉化的影响。 

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

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 ...

科研

医学

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 ...

Heterotopic Cervical Heart Transplantation in Mice

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

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

颈椎异位心脏移植小鼠

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

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

肝分型与门静脉结扎大鼠分期肝切除 (ALPPS) 手术模型的建立

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

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. ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

肝硬化大鼠门脉高压综合征的侵袭性血流动力学特征

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

通过结合70%部分切除术和对乙酰氨基酚，生成急性肝衰竭大鼠模型

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

Minimizing Post-Infusion Portal Vein Bleeding during Intrahepatic Islet Transplantation in Mice

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the kidney capsule in most rodent preclinical islet transplantation studies. This model is commonly used because murine intrahepatic islet transplantation is technically challenging, and a high percentage of mice could die from surgical complications, especially bleeding from the injection site post-transplantation. In this study, two procedures that can minimize the incidence of post-infusion portal vein bleeding are demonstrated. The first method applies an absorbable hemostatic gelatin sponge to the injection site, and the second method involves penetrating the islet injection needle through the fat tissue first and then into the portal vein by using the fat tissue as a physical barrier to stop bleeding. Both methods could effectively prevent bleeding-induced mouse death. The whole liver section showing islet distribution and evidence of islet thrombosis post-transplantation, a typical feature for intrahepatic islet transplantation, were presented. These improved protocols refine the intrahepatic islet transplantation procedures and may help laboratories set up the procedure to study islet survival and function in pre-clinical settings.

Here we present refined surgical procedures on successfully performing intraportal islet transplantation, a clinically relevant but technically challenging surgical procedure, in mice.

尽量减少小鼠肝内胰岛移植期间输注后门静脉出血

Although the liver is currently accepted as the primary transplantation site for human islets in clinical settings, islets are transplanted under the ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the number of heart failure patients waiting for transplantation is still increasing. The normothermic ex situ preservation technique has been established as a comparable method to the conventional static cold storage technique. The main advantage of this technique is that donor hearts can be preserved for up to 12 h in a physiologic condition. Moreover, this technique allows resuscitation of the donor hearts after circulatory death and applies required pharmacologic interventions to improve donor function after implantation. Numerous animal models have been established to improve normothermic ex situ preservation techniques and eliminate preservation-related complications. Although large animal models are easy to handle compared to small animal models, it is costly and challenging. We present a rat model of normothermic ex situ donor heart preservation followed by heterotopic abdominal transplantation. This model is relatively cheap and can be accomplished by a single experimenter.

Here, we present an assessment protocol of a heterotopically implanted heart after normothermic ex situ preservation in the rat model.

异位灌注异位心脏移植大鼠模型

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the ...

体内心脏移植研究内皮到间充质转化

A Neonatal Heterotopic Rat Heart Transplantation Model for the Study of Endothelial-to-Mesenchymal Transition

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and precludes patients with congenital critical aortic stenosis and hypoplastic left heart syndrome (HLHS) from curative anatomical biventricular surgical repair. Surgical resection is currently the only available therapeutic option, but EFE often recurs, sometimes with an even more infiltrative growth pattern into the adjacent myocardium.
To better understand the underlying mechanisms of EFE and to explore therapeutic strategies, an animal model suitable for preclinical testing was developed. The animal model takes into consideration that EFE is a disease of the immature heart and is associated with flow disturbances, as supported by clinical observations. Thus, the heterotopic heart transplantation of neonatal rat donor hearts is the basis for this model.
A neonatal rat heart is transplanted into an adolescent rat&#39;s abdomen and connected to the recipient&#39;s infrarenal aorta and inferior vena cava. While perfusion of the coronary arteries preserves the viability of the donor heart, flow stagnation within the LV induces EFE growth in the very immature heart. The underlying mechanism of EFE formation is the transition of endocardial endothelial cells to mesenchymal cells (EndMT), which is a well-described mechanism of early embryonic development of the valves and septa but also the leading cause of fibrosis in heart failure. EFE formation can be macroscopically observed within days after transplantation. Transabdominal echocardiography is used to monitor the graft viability, contractility, and the patency of the anastomoses. Following euthanasia, the EFE tissue is harvested, and it shows the same histopathological characteristics as human EFE tissue from HLHS patients.
This in vivo model allows for studying the mechanisms of EFE development in the heart and testing treatment options to prevent this pathological tissue formation and provides the opportunity for a more generalized examination of EndMT-induced fibrosis.

作者聚焦：用于研究内皮-间充质转化的新生儿异位大鼠心脏移植模型  

This work presents an animal model of endothelial-to-mesenchymal transition-induced fibrosis, as seen in congenital cardiac defects such as critical aortic stenosis or hypoplastic left heart syndrome, which allows for detailed histological tissue evaluation, the identification of regulatory signaling pathways, and the testing of treatment options.

新生儿异位大鼠心脏移植模型在内皮间充质转化研究中的应用

Endocardial fibroelastosis (EFE), defined by subendocardial tissue accumulation, has major impacts on the development of the left ventricle (LV) and ...