Este estudio fue financiado por la &quot;Deutsche Forschungsgemeinschaft&quot; (DFG). K. Schleimer fue apoyada por el Lise Meitner de Becas del Ministerio de Ciencia e Investigación de NRW. Nos gustaría dar las gracias a la señora Mary-Joan Blümich por su asistencia en la edición excelente. Dr. Maria Kokozidou se financió con Fresenius 2011_A61.

1. Animales Experimentales <ol><li> Obtenga una aprobación del Comité de Ética local para todos los experimentos que se llevarán a cabo, de conformidad con las directrices de protección animal FELASA (K 40, 17/99). </li><li> Utilice ratas Lewis macho disponible para comprar desde cualquier criador comercial (por ejemplo, Charles River Wiga GmbH, Sulzfeld, Alemania). En este caso, se utilizaron 224 animales (112 donantes, 112 destinatarios) para realizar 112 trasplantes. Los receptores fueron sacrificados a las 6 semanas después de la operación. En el momento de la operación, los animales deben pesar 250 a 350 g. </li><li> Mantener a los animales en las habitaciones climatizadas con acceso libre a comida y agua. </li></ol> 2. Anestesia y Preparación Operativo <ol><li> Coloque la rata en una caja de plexiglás conectado a la máquina de anestesia con una tasa de flujo de oxígeno de 1 L / min, y un vaporizador con gas anestésico isofluorano al 5% para inducir la anestesia. Mantener la anestesia en el 1,5% isofluorano. </li><li> Coloque las ratas endecúbito supino sobre una placa de calentamiento. </li><li> Retire la piel del abdomen con una crema de depilación. </li><li> Preparar un campo quirúrgico estéril de la siguiente manera: Utilice un desinfectante para limpiar la mesa de trabajo. Para la desinfección del sitio de la incisión el uso de clorhexidina primero, segundo alcohol y, finalmente, una solución exfoliante (por ejemplo, clorhexidina y 70% de alcohol isopropílico). Póngase una bata quirúrgica desechable y guantes quirúrgicos. </li><li> Utilizar instrumentos de microcirugía estériles. </li><li> Utilice un microscopio quirúrgico en el porcentaje de 12 a 20 veces. </li></ol> 3. Procedimiento Quirúrgico <ol><li> Donantes de operación Preparación <ol><li> Lleve a cabo una laparotomía media y resección del lóbulo lateral izquierdo y el lóbulo medio del injerto según lo descrito por Higgins. 14 </li><li> A continuación, preparar el ligamento hepatoduodenal. En primer ligar el conducto biliar para inducir congestión y dilatación de la pequeña conducto biliar. Entonces ligar y seccionar el gastroduodevena y arteria nal. Después, aislar el tronco celíaco y la arteria hepática común y seccionar la arteria gástrica izquierda y la arteria esplénica. Incisión en el conducto biliar congestionada, e introducir un stent 20-G en él. El posicionamiento de la endoprótesis debe garantizar el drenaje de todo el sistema de conductos biliares hiliar. Seccionar el conducto biliar distal al stent. </li><li> Aislar el segmento infrahepática de la vena cava derecha por encima de la vena renal derecha. Perfusión </li><li> Después de la ligadura de la aorta por encima del tronco celíaco, sujetar la vena porta y incisión ella. Introducir el catéter de perfusión (14 G) en la vena porta, y realizar la perfusión con 30 ml de-triptófano-cetoglutarato histidina (HTK) solución al (12 cm H2O, 4 ° C). </li><li> Cortar el diafragma y la vena cava suprahepática para permitir que el líquido de perfusión fluya hacia fuera. </li><li> Cortar el tronco celíaco, junto con un parche aórtico y enjuague con solución de histidina-triptófano-cetoglutarato. Explantación </li><li> Seccionar la vena cava infrahepática justo encima de la vena renal derecha. </li><li> Luego, colocar un stent con un diámetro de 0,3 mm en la vena porta y fijarlo con una ligadura. Durante la siguiente operación, la vena porta se arterializada través de la arteria renal derecha. Seccionar la vena porta distal al stent. </li><li> Después de la ligadura de la vena cava suprahepática, seccionar junto con un manguito de diafragma. </li><li> Explante el injerto y almacenarlo en histidina-triptófano-cetoglutarato (HTK) solución al a 4 ° C en una bolsa de plástico con hielo. Suturas Lugar de esquina (8-0 Ethilon) en la vena cava infrahepática. </li></ol></li><li> Operación Receptor <ol><li> Lleve a cabo una laparotomía media y preparar la vena cava infrahepática. Entonces aislar la arteria renal derecha y el uréter, ligar el uréter y seccionar ella. Sujete la arteria renal derecha en su origen, a continuación, ligar la vena renal derecha. Luego, después de la transección de ambos revasos nales, realizan la nefrectomía. Posteriormente, acortar la arteria renal derecha, enjuague con solución salina de heparina (40 UI de heparina solución salina / ml). </li><li> Seccionar la vena renal derecha proximal a la ligadura después de sujeción proximal y distal de la vena cava infrahepática para crear un lumen ovales en la vena cava infrahepática. Enjuague la vena cava con solución salina de heparina. Implante el injerto en la región renal derecha del abdomen del destinatario. </li><li> Anastomose infrahepática extremo a lado de la vena cava del donante a la recipient`s vena cava con una sutura continua 8-0 Ethilon. </li><li> Después de aclarar con solución de heparina / solución salina, introducir el stent en la vena porta en la arteria renal derecha (Figura 3). Conecte la vena porta y la arteria renal con 2 puntos (8-0 Ethilon). Fijar el stent que une en la arteria renal con una ligadura (trenzado de seda negro 7-0). </li><li> Para permitir la reperfusión, primero retire la pinza arterial, entonces ambosabrazaderas venosos (Figura 4). </li><li> Preparar la aorta infrarrenal, luego coloque una proximal y distal y microclamp incidir la aorta justo entre las dos pinzas. Lleve a cabo una anastomosis término-lateral entre la aorta y el tronco celíaco del donante, el uso de un parche de aorta, con una sutura continua 10-0 Ethilon. </li><li> Inserte el conducto biliar en el duodeno: Aplicar una sutura en bolsa de tabaco en la pared duodenal, hacer una pequeña incisión en la pared duodenal justo en el medio de la bolsa de tabaco. Introducir el conducto biliar con stent en el duodeno, y apretar la sutura en bolsa de tabaco. Finalmente, mantenga el duodeno y el conducto biliar firmemente juntos con dos puntos de sutura (Ethilon 8-0) (Figura 5). </li><li> A continuación, realice una resección subtotal del hígado nativo, descrito por primera vez por Emond et al. para imitar la insuficiencia hepática aguda. 7 </li><li> Fijar la vena cava suprahepática del injerto, junto con un manguito de diafragma, a la pared abdominal lateral de la reincipiente con 4-0 Vicryl. Enjuague el abdomen con solución salina (Figura 6). </li><li> Para la capa muscular de usar una sutura continua de Vicryl 4-0 y para la cutánea una sutura continua de Vicryl 3-0. </li></ol></li></ol> 4. Recuperación y Manejo Postoperatorio <ol><li> Deje que los animales se recuperen de la anestesia bajo una lámpara infrarroja calentamiento. </li><li> Inyectar buprenorfina a una dosis de 0,03-0,05 mg / kg BW sc cada 8-12 horas durante 2 días. </li><li> Examine los animales en relación con el estado general y signos de enfermedad o infecciones. </li></ol> 5. Cálculo de la inicial Nativo Peso Hígado Calcular el peso inicial hígado nativo de la siguiente manera: restar el peso de los lóbulos del hígado resecados del peso del hígado completo (2,66% del peso corporal (este valor con una desviación estándar de 0,49% +/- se determinó en experimentos anteriores con n = 80 animales de la misma cepa y unge)). 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. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterialization+of+the+portal+vein+in+orthotopic+and+auxiliary+liver+transplantation.">Arterialization of the portal vein in orthotopic and auxiliary liver transplantation.</a> Transplantation. 60, 877-879 (1995).</li><li>Erhard, 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=Auxiliary+liver+transplantation+with+arterialization+of+the+portal+vein+for+acute+hepatic+failure.">Auxiliary liver transplantation with arterialization of the portal vein for acute hepatic failure.</a> Transpl Int. 11, 266-271 (1998).</li><li>Emond, J., Capron-Laudereau, M., Meriggi, F., Bernuau, J., Reynes, M., Houssin, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extent+of+Hepatectomy+in+the+rat.">Extent of Hepatectomy in the rat.</a> Eur Surg Res. 21, 251-259 (1989).</li><li>Mas, A., Rode`s, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fulminant+hepatic+failure.">Fulminant hepatic failure.</a> Lancet. 349, 1081-1085 (1997).</li><li>Terpstra, O. 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>

Todos los autores declaran intereses en conflicto.

La primera heterotópico auxiliar trasplante de hígado de rata fue descrito por Lee et al., 16 en 1966 seguido por Hess. 17 La vena porta del injerto se anastomosa con la vena porta del receptor. Durante este procedimiento, se ve comprometido el suministro de sangre venosa portal al hígado nativo. En 1991 Hong realiza HALT con PVA a través de la arteria ilíaca del destinatario. 13 Sin regulación del flujo sanguíneo portal masiva y se produjo co...

Insuficiencia hepática aguda plantea un problema grave en la medicina clínica que lleva a una tasa de mortalidad del 60-80%. 8 En la insuficiencia hepática aguda, el trasplante hepático auxiliar es una terapia alternativa prometedora para superar la fase aguda. El hígado nativo se deja de regenerarse y siguiendo a la regeneración de los medicamentos inmunosupresores pueden ser descontinuados. Como consecuencia, el injerto se convertirá atrófica. Sin embargo aún se discute la técnica quirúrgica de trasplante hepático auxiliar. 1.3 En este contexto, Terpstra informó la falta de espacio para el injerto, la sangre portal robar fenómeno y el problema de la congestión venosa y drenaje. 9-10 vena porta del donante fue anastomosada extremo a un lado a la vena porta del receptor. Esto dio lugar a la competencia flujo portal entre el injerto y el hígado nativo y en la trombosis de la vena porta. La mayoría de los problemas técnicos se pueden evitar con el uso del método APOLT (orthotopic implantación de segmentos auxiliares). 4 Sin embargo, este método requiere resecciones extensas tanto del hígado nativo y el. injerto 4 En 1998, Erhard y su equipo desarrollaron el trasplante heterotópico auxiliar hígado (HALT) utilizando arterialización de la vena porta (PVA) (Figura 1 ). El hilio y la vena porta del hígado nativo se respetan con el uso de este método. Esto ahorra el destinatario de los efectos secundarios no deseados, como la trombosis de la vena porta y de la sangre portal fenómeno de robo, lo que podría perturbar la recuperación del hígado nativo. Además, no se requieren extensas resecciones hepáticas del hígado nativo y del injerto para este método, por consiguiente, con menos sangrado y más masa total del parénquima hepático restante para puentear (injerto) y la regeneración (hígado nativo). Esta técnica mostró prometedores resultados clínicos iniciales. 5-6 El objetivo de este trabajo experimental fue el desarrollo de una técnica HALT con arterialización vena porta enla rata para fines de investigación (Figura 2). En los experimentos preliminares no tomamos en cuenta la regulación del flujo sanguíneo de la vena portal arterializada, que dio lugar a la congestión del injerto trasplantado después de la reperfusión y sangrado masivo de la cápsula hepática. 11 Desde entonces, el flujo de sangre en la vena porta arterializada se reguló en un stent con un diámetro interior de 0,3 mm y una longitud de 8 mm. Experimentos piloto había demostrado que un diámetro del stent de 0,3 mm produce un flujo promedio de sangre en la vena porta arterializada que está dentro del intervalo fisiológico superior (1.7 +/- 0,4 ml / min / g de peso de hígado). 12 Nuestros resultados sin regulación del flujo sanguíneo correlacionar con los resultados de Hong, que realizó HALT con PVA a través de la arteria ilíaca del destinatario. 13 En sus experimentos sin regulación del flujo sanguíneo portal masiva y se produjo congestiones sinusoide. El objetivo de nuestro video es demonstrate paso a paso esta técnica quirúrgica experimental ne.w en la rata. A largo plazo, la técnica propuesta puede ser utilizada en los estudios sobre la regeneración del hígado y de la competencia inter-hígado y arterialización vena porta, hiperperfusión vena porta y la regulación del flujo sanguíneo portal.

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<tbody>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Braided Silk black 7-0&#160;</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">892100</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 3-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J285G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Vicryl 4-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">J284G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 8-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">1714G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:216px;">Ethilon 10-0</td>
			<td style="width:71px;">Ethicon Products</td>
			<td style="width:119px;">7770G</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 14G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758853</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Vasofix safety 20G</td>
			<td style="width:71px;">B Braun</td>
			<td style="width:119px;">2758818</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="201">
			<td height="201" style="height:201px;width:216px;">2 French (.012&#8221;/0.3mm ID x .025&#8221;/0.6mm OD) x 24&#8221;/60cm polyurethane catheter</td>
			<td style="width:71px;">Access technologies a division of Norfolk Medical, Inc., Illinois, USA</td>
			<td style="width:119px;">CNC-2P</td>
			<td style="width:167px;"></td>
		</tr>
		<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>
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heterotopic auxiliary rat liver transplantation with flow-regulated portal vein arterialization in acute hepatic failure

En la insuficiencia hepática aguda del trasplante hepático auxiliar es un enfoque alternativo interesante. El objetivo es proporcionar un soporte temporal hasta que el hígado nativo no se ha regenerado. 1-3 El APOLT-método, el implante ortotópico de segmentos- auxiliar evita la mayoría de los problemas técnicos. Sin embargo este método requiere de resecciones extensas tanto en el hígado nativo y el. Injerto 4 En 1998, Erhard desarrolló el trasplante heterotópico auxiliar hígado (HALT) utilizando arterialización de la vena porta (PVA) (Figura 1). Esta técnica mostró prometedores resultados clínicos iniciales. 5-6 Hemos desarrollado una técnica HALT-con-regulada flujo de PVA en la rata para examinar la influencia de PVA-flujo regulado en la morfología y función (Figura 2) injerto. Un injerto de hígado reducido a un 30% de su tamaño original, se implantó heterotópicamente en la región renal derecha del receptor después de la explantación del riñón derecho.60; La vena cava infra-hepática del injerto se anastomosa con la vena cava infrahepática del destinatario. El arterialización de la vena porta del donante se llevó a cabo a través de la arteria renal derecha del destinatario con la técnica de stent. La regulación del flujo sanguíneo de la vena porta arterializada se logró con el uso de un stent con un diámetro interno de 0,3 mm. El tronco celíaco del injerto fue implantado era de extremo a lado anastomosa con la aorta del receptor y el conducto biliar en el duodeno. Una resección subtotal del hígado nativo se realizó para inducir insuficiencia hepática aguda. 7 De esta manera se realizaron 112 trasplantes. La tasa de supervivencia perioperatoria fue del 90% y la tasa de supervivencia de 6 semanas fue del 80%. Seis semanas después de la operación, el hígado regenerado nativo, mostrando un aumento de peso de 2,3 ± 0,8 g a 9,8 ± 1 g. En este momento, el peso del injerto disminuyó de 3,3 ± 0,8 g a 2,3 ± 0,8 g. </p&gt; Hemos sido capaces de obtener resultados prometedores a largo plazo en términos de morfología y función del injerto. HALT con flujo regulado PVA tiende un puente fiable insuficiencia hepática aguda hasta que regenera el hígado nativo.

La duración media del trasplante fue de alrededor de 150 min. El tiempo de isquemia fría fue de 64 +/- 7 min, el tiempo de isquemia caliente fue de menos de 25 min. La tasa de supervivencia perioperatoria fue del 90% y la tasa de supervivencia de 6 semanas fue del 80%. La pérdida de 20% se debió a una peritonitis debido a la dislocación del stent y la fuga biliar (cuatro animales) durante la primera semana postoperatoria y debido a la neumonía, absceso intraabdominal e íleo (siete ani...

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

Trasplante hepático auxiliar proporciona un soporte temporal en la insuficiencia hepática aguda, hasta que la regeneración del hígado no. El trasplante heterotópico auxiliar hígado (HALT) con arterialización vena porta (PVA) hace una función hepática suficiente. Hemos desarrollado una técnica análoga en la rata, para examinar la influencia de la arterialización de la vena porta en la morfología y la función del injerto.

Heterotópico Rata auxiliar Trasplante de Hígado Con Flujo regulado Portal Vein Arterialización en aguda Insuficiencia Hepática

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 vistas.  University Hospital RWTH Aachen.  Trasplante hepático auxiliar proporciona un soporte temporal en la insuficiencia hepática aguda, hasta que la regeneración del hígado no. El trasplante heterotópico auxiliar hígado (HALT) con arterialización vena porta (PVA) hace una función hepática suficiente. Hemos desarrollado una técnica análoga en la rata, para examinar la influencia de la arterialización de la vena porta en la morfología y la función del injerto. 

Video: 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.

Procedimientos Quirúrgicos para un modelo de rata de trasplante hepático ortotópico parcial con reconstrucción arterial hepática

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Investigación

Medicina

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.

Rata heterotópico abdominal Corazón / Trasplante de un solo pulmón en una configuración de volumen cargado

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.

Heterotópico cervical trasplante cardiaco en ratones

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.

Modelo de la rata de la Asociación de partición hepática y ligadura de la vena porta para procedimiento efectuado hepatectomía (ALPPS)

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.

Volver a arterializada rata parcial trasplante del hígado con una en vivo orientada al barco 70% hepatectomía

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.

Caracterización hemodinámica invasiva del síndrome de hipertensión Portal en cirróticos ratas

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.

Generación de un modelo de rata de insuficiencia hepática aguda mediante la combinación de 70% de hepatectomía parcial y paracetamol

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.

Minimizar el sangrado de la vena porta posterior a la infusión durante el trasplante intrahepático de islotes en ratones

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.

Modelo de rata de trasplante cardíaco heterotópico con perfusión ex-situ normotémica

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

Trasplante de corazón in vivo para estudiar la transición endotelial a mesenquimal

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.

Autor destacado: Un modelo de trasplante de corazón de rata heterotópico neonatal para el estudio de la transición endotelial a mesenquimal  

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.

Un modelo de trasplante de corazón de rata heterotópico neonatal para el estudio de la transición endotelial a mesenquimal

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