TC fue apoyada por la Swiss National Science Foundation (SCORE concesión 3232230 a 126233). El estudio fue apoyado por la Fundación Artères, la Europa de Astellas y la Fundación Boninchi.

1. Operación de los donantes <ol><li> Anestesie ratas macho que pesaban 200 ± 20 gramos con isoflurano en el cono de la máscara (3% para la inducción, el 2% durante la operación, un flujo de aire l / min, FiO2 70%). </li><li> Realice una laparotomía mediana y gran transversal y cauterizar los vasos epigástricos en ambos lados. </li><li> Cortar el ligamento falciforme, y separar la vena diafragmática izquierda de la vena cava suprehepatic. Por último, se divide la vena diafragmática entre dos ligaduras de seda 7-0. </li><li> Cortar el triangular de la izquierda y los ligamentos gastro-hepáticos. </li><li> Divida el ligamento hepato-esofágico y la arteria entre el 7-0 ligaduras (coagulación es una alternativa válida). </li><li> Aislar el infrahepática la vena cava hasta la vena renal izquierda. Separe la vena renal derecha de los tejidos circundantes y se divide entre un 10-0 y una ligadura de 7-0, en sus extremos proximal y distal, respectivamente. </li><li> Divida la vena suprarrenal derecha entre el 7-0 ligatures libre y el hígado de sus ligamentos posteriores al cortar en una suave tracción. </li><li> Aislar la vena gastrosplénico y se divide entre un 10-0 y una ligadura 7-0, en sus extremos proximal y distal, respectivamente. </li><li> Divida la arteria hepática adecuada entre 7-0 relaciones. </li><li> Aislar la vena duodeno-páncreas y lo dividió entre el 10-0 y una ligadura de 7-0, en su extremo proximal y distal, respectivamente. </li><li> Inserte un 22G, 3,5 mm del stent en el conducto biliar común, practicando una pequeña incisión en la distancia de 1 cm desde el hilio hepático. Asegure el stent en la posición con una ligadura de 7-0. </li><li> Inyectar 10 UI de heparina, se diluyó en 1 ml de solución salina normal, a través de la vena dorsal del pene. </li><li> Canular la vena porta (la medida de lo posible, desde el hilio) con una aguja 21G y lave cuidadosamente el hígado con 20 ml de lactato de Ringer fría (o con otras soluciones de preservación). Al mismo tiempo, cortar la vena cava por debajo de las venas renales para permitir una adecuada outflow. La perfusión del hígado en frío no debe durar entre uno y dos minutos. </li><li> Cortar la vena porta debajo de la vena duodeno-pancreático. </li><li> Cortar el conducto biliar común distalmente a la ligadura alrededor del stent. </li><li> Completa la incisión de la vena cava infrahepática. </li><li> Cortar la vena cava suprahepática descremada al diafragma y eliminar el hígado. </li><li> Coloque el hígado en un recipiente lleno de lactato de Ringer frío y laicos de la cuenca en una plataforma de hielo. </li></ol> 2. Preparación del injerto <ol><li> Insertar la vena porta en su manguito, a continuación, volver el recipiente alrededor del borde para exponer la íntima. Asegure el segmento evertido en el manguito con una ligadura de 7-0. </li><li> Repita la misma operación en el infrahepática la vena cava. </li><li> Coloque una abrazadera de micro-(4-6 mm de longitud) en la parte proximal de la infrahepática la vena cava. Esta operación evita la pérdida de sangre después de la reperfusión del portal. </li><li> Revocar el hígado e insertar dos puntos de sutura prolene 8-0en los bordes laterales opuestos de la vena cava suprahepática, de afuera hacia adentro. </li><li> Guarde el injerto (sumergido en solución de Ringer lactato) en el refrigerador a 0-4 ° C. </li></ol> 3. Operación del destinatario <ol><li> Anestesie ratas macho que pesaban 200 ± 20 gramos con isoflurano en el cono de la máscara (3% para la inducción, el 2% durante la operación, un flujo de aire l / min, FiO2% 70) y seguir una almohadilla caliente. Es de destacar que el peso receptores y donantes deben coincidir (± 40 gramos). </li><li> Inyectar 10 ml de solución salina normal por vía subcutánea y 0,03 g de piperacilina / tazobactam por vía intramuscular antes de la laparotomía. </li><li> Realice una línea media xipho-púbica laparotomía. </li><li> Invierta el proceso xifoides con una pinza de auto-estáticos con el fin de exponer mejor el hígado. </li><li> Corte los ligamentos anteriores y dividir la vena diafragmática izquierda entre dos relaciones 7-0. </li><li> Divida el ligamento hepato-esofágico y la arteria entre las ligaduras (7-0 coagulation es una alternativa válida). No se muestra en el vídeo (idéntico al funcionamiento de los donantes). </li><li> Aislar el infrahepática la vena cava hasta la vena renal derecha. </li><li> Divida la vena suprarrenal derecha entre el 7-0 ligaduras y sin el hígado de sus ligamentos posteriores al cortar en una suave tracción (no se muestra en el vídeo (idéntico al funcionamiento de los donantes)). </li><li> Coloque una ligadura de 7-0 alrededor del conducto biliar común, justo debajo de su división. Dicho de otra ligadura de un milímetro por debajo y cortar en el medio. </li><li> Aislar la arteria hepática en el hilio y se divide entre el 7-0 ligaduras. </li><li> Suavemente separa las ramas portales izquierda y derecha. Preste atención para no dañar la arteria hepática común o la vena gastro-esplénico. </li><li> Coloque el anillo de la vena cava alrededor de la vena cava, cranealmente a la vena renal derecha. A continuación, fije la pared del vaso al anillo mediante la colocación de cuatro puntos cardinales 9-0 puntos de sutura. </li><li> Repetir la misma operación con el anillo en el portal de la vena porta. </li&gt; <li> Inyectar 10 UI de heparina, se diluyó en 1 ml de solución salina normal, a través de la vena dorsal del pene. </li><li> Establecer la anestesia en 0,25% y la FiO2 en 100%. </li><li> Fije el infrahepática la vena cava por encima de la vena renal derecha, y luego fijar la vena porta justo por encima de la vena gastro-esplénico. Por último apretar el vena cava suprahepática y asegúrese de incluir al menos 2 mm de diafragma. </li><li> Cortar la vena suprahepática cerca de cava para el parénquima. </li><li> Cortar la izquierda y las ramas portales derechas y dividir el tabique en el medio. </li><li> Cortar el infrahepática la vena cava cerca del parénquima, teniendo cuidado de no incidir cualquiera de las suturas previamente colocadas. Retirar el hígado y colocar el injerto en la cavidad abdominal. </li><li> Lleve a cabo una anastomosis término-terminal entre la vena cava suprahepática del injerto y del receptor. Comience con la sutura de la pared posterior de izquierda a derecha, a continuación, proceder en la pared anterior del derecho a la le-Mantenga pies de la sutura anterior suelta hasta el final con el fin de permitir que un lavado abundante de la luz interior antes de apretar los extremos. </li><li> Inserte la rama caudal de la aproximador en el mango del anillo del portal, a continuación, introduzca la vena porta del injerto en la hendidura de la rama craneal de la aproximador, con el brazalete del portal para encontrar su posición en la ranura. </li><li> Cerrar la aproximador al enjuagar el recipiente de la vena porta con solución salina normal y permitir que el manguito para insertar en el recipiente. Fije la vena porta del receptor alrededor de su puño con un empate 7-0 circunferencial. </li><li> Retire el aproximador. Abra la pinza en la vena cava suprahepática y la vena porta con el fin de: flujo portal está restablecido. </li><li> Retire el anillo del portal mediante la reducción de las ligaduras de 9-0, con extra-finas tijeras. </li><li> Inserte la rama caudal de la aproximador en el mango del anillo cava, a continuación, insertar el infrahepática la vena cava del injerto en la ranura del cráneo<html> rama del aproximador, con el manguito de la vena cava para encontrar su posición en la ranura. </li><li> Cierre el aproximador, durante la descarga del destinatario infrahepática la vena cava con solución salina normal y permitir que el manguito para insertar en el buque. Asegure el receptor la vena cava alrededor de su puño con un empate 7-0 circunferencial. </li><li> Retire el aproximador. Abra las microclamps en el infrahepática la vena cava del injerto y receptor, con el fin de: flujo venoso está restablecido. </li><li> Realice una incisión parcial del conducto del receptor biliar común, aplique una tracción suave hacia la cabeza, e insertar el lado libre del stent previamente insertado en el conducto biliar del injerto. Asegure el stent con una circunferencia 7-0 empate. </li><li> Cerca del animal por capas, con 5-0 puntos de sutura. Deje que el agua libre y comida de despertar. </li><li> Administrar analgesia eficaz de acuerdo a protocolos institucionales locales. Una duración mínima de 24 h se recomienda. </li></ol>_content &quot;&gt; De la nota: de extremo a extremo anastomosis de la arteria hepática puede llevar a cabo, cuando sea requerido por el diseño experimental (la arteria no es necesaria para la supervivencia a largo plazo después del trasplante de hígado de la rata). 4. Los resultados representativos La técnica descrita puede ser dominado con éxito por nuestro equipo después de aproximadamente 10 cirugías de entrenamiento. Permitió que el 100% a largo plazo (&gt; 21 días) la supervivencia en 36 consecutivos Lewis a Lewis y trasplantes oscuros Agouti a Lewis, después de que el corto periodo de formación (un número de receptores singénicos injerto hepático se han mantenido vivas durante más de 12 Desde hace meses). Post-trasplante syngeneic día 1 pruebas de función hepática mostraron aspartato transaminasa mediana (AST): 84 U / l (51,7 a 92,7) y la mediana de la alanina aminotransferasa (ALT): 172,5 U / l (127,5-240,2). La fase de anhepática medio (desde la vena porta de sujeción a la reperfusión del injerto) fue de 14 ± 2 min, con el paso de ser el más largo de la vena cava suprahepática anastomosis (s corriendoFUTURO) Tiempo de 9 ± 2 min. El sistema Quick-enlazador permitido el posicionamiento de 1,55 mm de diámetro en la vena porta esposas y 2,40 mm de diámetro puños en la vena cava (receptores de peso de 200 ± 20 g). Por el contrario, cuando tratamos de transplante usando la técnica de Kamada, que no podían exceder de 1,40 mm y 2,16 mm, respectivamente (las ratas de contrapartida para la tensión y el peso). Como se muestra en la Figura 7, rápida-enlazador anillos están diseñados para mantener los vasos receptores en óptimo de estiramiento, permitiendo mínimo pinza y el desgaste de longitud, lo que resulta en el más cercano a fisiológicas resultados hemodinámicos. <img src="/files/ftp_upload/4143/4143fig1.jpg" alt="Figura 1" /> Figura 1. Medidas Anillos. <img src="/files/ftp_upload/4143/4143fig2.jpg" alt="Figura 2" /> Figura 2. Medidas Microclamps. <img src="/files/ftp_upload/4143/4143fig3.jpg" alt="Figura 3" /> Figura 3. Quick-enlazador kit. <img src="/files/ftp_upload/4143/4143fig4.jpg" alt="Figura 4" /> Figura 4. Rápido enlazador armado. <img src="/files/ftp_upload/4143/4143fig5.jpg" alt="Figura 5" /> Figura 5. Rápido enlazador cerrado. <img src="/files/ftp_upload/4143/4143fig6.jpg" alt="Figura 6" /> Figura 6. Abrazadera SHVC. <img src="/files/ftp_upload/4143/4143fig7.jpg" alt="Figura 7" /> Figura 7. Classic vs rápida enlazador asistidos por anastomosis del manguito-hemodinámica. <img src="/files/ftp_upload/4143/4143fig8.jpg" alt="Figura 8" /> Figura 8. De los donantes y la preparación de la piel del destinatario y la incisión.

<ol>
	<li>Starzl, T. E., Marchioro, T. L., Porter, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+and+clinical+observation+after+homotransplantation+of+the+whole+liver.">Experimental and clinical observation after homotransplantation of the whole liver.</a> Revue internationale d'hepatologie. 15, 1447 (1965).</li><li>Starzl, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+liver+transplantation.">Evolution of liver transplantation.</a> Hepatology. 2, 614S-636S (1982).</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 (1979).</li><li>Kashfi, A. <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> Transplantation proceedings. 37, 185-188 (2005).</li><li>Hori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+and+innovative+techniques+for+liver+transplantation+in+rats:+A+surgical+guide.">Comprehensive and innovative techniques for liver transplantation in rats: A surgical guide.</a> World Journal of Gastroenterology: WJG. 16, 3120 (2010).</li><li>Oldani, G. <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+technique+for+rat+liver+transplantation+using+Quick+Linker+system:+a+preliminary+result.">A novel technique for rat liver transplantation using Quick Linker system: a preliminary result.</a> Journal of Surgical Research. 149, 303-309 (2008).</li><li>Nosaka, T., Bowers, J. L., Cay, O., Clouse, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biliary+complications+after+orthotopic+liver+transplantation+in+rats.">Biliary complications after orthotopic liver transplantation in rats.</a> Surg. Today. 29, 963-965 (1999).</li><li>Li, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+incidence+of+biliary+complications+in+rat+liver+transplantation:+can+we+avoid+it.">High incidence of biliary complications in rat liver transplantation: can we avoid it.</a> World J. Gastroenterol. 17, 3140-3144 (2011).</li><li>Peng, Y., Gong, J. P., Yan, L. N., Li, S. B., Li, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+two-cuff+technique+for+orthotopic+liver+transplantation+in+rat.">Improved two-cuff technique for orthotopic liver transplantation in rat.</a> Hepatobiliary Pancreat. Dis. Int. 3, 33-37 (2004).</li><li>Urata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+survival+in+rat+liver+transplantation+with+extended+cold+preservation:+role+of+portal+vein+clamping+time.">Decreased survival in rat liver transplantation with extended cold preservation: role of portal vein clamping time.</a> Hepatology. 28, 366-373 (1998).</li></ol>

No hay conflictos de interés declarado.

El hígado de rata actual modelo de trasplante puede establecerse fácilmente. En nuestra experiencia, algunos puntos clave que ayudan a minimizar el riesgo de muerte (≤ 20%). Las muertes tempranas, producidas en las tres primeras horas de la reperfusión, son más a menudo debido a un infarto intestinal, como consecuencia de un portal de tiempo de fijación. Por tanto, se recomienda mantener la fase anhepática en el límite ideal de 15 minutos (20 minutos todavía aceptable). Central de la embolia de aire también puede ser una causa de muerte súbita y es obligatorio para eliminar todas las venas antes de la anastomosis. Las muertes que se producen entre el día postoperatorio y tres son a menudo debido a la insuficiencia hepática o la trombosis de la vena cava. Uno por lo tanto, deben evitar cualquier injerto agresiva lavado antes de la explantación en el donante (que lavar con 20 ml durante aproximadamente 60 segundos) y garantizar una conservación adecuada en frío (inmersión total en 0-4 ° C solución de Ringer lactato). El riesgo de trombosisse puede disminuir reduciendo al mínimo la manipulación de la íntima y preservar su integridad. No se recomienda el uso de la anticoagulación después de la cirugía. Las muertes entre los días cuatro y nueve postoperatorio se relaciona a menudo con colangitis bacteriana 7. Basándonos en nuestra experiencia, una sola dosis de piperacilline / tazobactam (0,03 g) dada antes de la laparotomía es suficiente para reducir al mínimo el riesgo de infección del conducto biliar común. La muerte es un evento raro después del día 10. Sin embargo, debido a la ausencia de flujo arterial, algunos problemas de conductos biliares se puede observar con cholastasis 8. Dependiendo de las necesidades del investigador una arterialización puede considerarse 9. De nota, un tiempo de isquemia fría de hasta 24 horas pueden ser considerados como aceptables 10. El sistema Quick-enlazador fue originalmente diseñado para llevar a cabo el trasplante de hígado con tres anastomosis del manguito (fase anhepática muy corto). Sin embargo, el TE tres manguitochnique requiere el uso de pinzas muy pequeñas para la vena cava suprahepática (SHVC) del manguito y que no ha sido ampliamente aceptado debido al riesgo de problemas de flujo de salida del hígado 11-14. Por lo tanto, favorecen un rápido enlazador asistida de tres manguito técnica sólo cuando los tiempos de isquemia caliente de menos de diez minutos se requieren. En la mayoría de los estudios sin embargo, las fases de implantación de hasta 15-18 minutos son aceptables, permitiendo el uso de la técnica de dos manguito, y un drenaje SHVC más fisiológico. Como se muestra en este documento de vídeo, de manera rutinaria utilizar el sistema rápido enlazador en la segunda parte de la implantación solamente. Mientras que algunos tiempo quirúrgico adicional es necesario para colocar las asas rápido enlazador, esta parte del procedimiento se asocia con riesgos mínimos y permite un acortamiento de la fase crítica, gracias a anhepática más fácil la inserción en los puños. Otras ventajas del sistema rápido enlazador son un mejor injerto-receptor alineación buque y el uso de amplios puños con mejores resultados hemodinámicos en comparación con eltécnicas descritas previamente (incluyendo el descrito por Kamada y col.) 6 (Fig. 7).

<table border="1"><tbody><tr><td> Herramienta / Reactivo </td><td> Empresa </td><td> Catálogo n. </td><td> Comentario </td></tr><tr><td> Portal del manguito </td><td> <a href="http://altecweb.com" target="_blank">altecweb.com</a> o <a href="http://smiths-medical.com" target="_blank">herreros medical.com-</a> </td><td> Bobina 01-96-1729 800/100/420 </td><td> Diámetro: 1.55mm Pared: 0.30mm Longitud: 3,5 mm </td></tr><tr><td> Infrahepática v cava manguito </td><td> <a href="http://altecweb.com" target="_blank">altecweb.com</a> o <a href="http://smiths-medical.com" target="_blank">herreros medical.com-</a> </td><td> Bobina 01-96-1733 800/100/540 </td><td> Diámetro: 2.40mm Pared: 0.30mm <br/&gt; Longitud: 3,5 mm </td></tr><tr><td> Stent del conducto biliar común </td><td> común venflon 22G </td><td></td><td></td></tr><tr><td> 7-0 corbatas de seda </td><td> <a href="http://georgetiemann.com" target="_blank">georgetiemann.com</a> </td><td> 160-1226-7 / 0 </td><td></td></tr><tr><td> 10-0 prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> 9-0 Prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> 8-0 Prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> Pinzas rectas micro </td><td> <a href="http://s-and-t.net" target="_blank">-s y-t</a>. Net </td><td> FRC-15RM-8 </td><td></td></tr><tr><td> Pinzas de micro curvas </td><td> <a href="http://s-and-t.net" target="_blank">s-y-t.net</a> </td><td> FRS-15-RM-8 </td><td></td></tr><tr><td> Disección de porta </td><td> <a href="http://s-and-t.net" target="_blank">s-y-t.net</a> </td><td> B-15-8.2 </td><td></td></tr><tr><td> Anastomosis porta </td><td> <a href="http://rumex.net" target="_blank">rumex.net</a> </td><td> 8-021T </td><td></td></tr><tr><td> Micro pinzas </td><td> <a href="http://finescience.com" target="_blank">finescience.com</a> </td><td> 18055-03 </td><td> 6 x 1 mm </td></tr><tr><td> Abrazaderas aplicador Micro </td><td> <a href="http://finescience.com" target="_blank">finescience.com</a> </td><td> 18057-14 </td><td></td></tr&gt; <tr><td> V cava suprahepática pinza </td><td> <a href="http://pharmap.ch" target="_blank">pharmap.ch</a> </td><td> 13.349.14 </td><td></td></tr><tr><td> Quick-Linker anillos </td><td> <a href="http://dltchiropody.co.uk" target="_blank">dltchiropody.co.uk</a> </td><td> PAR 10 </td><td> modificados hojas de bisturí </td></tr><tr><td> Quick-Linker aproximador </td><td> <a href="http://pharmap.ch" target="_blank">pharmap.ch</a> </td><td> UD-04-275 </td><td> pinza Kocher modificado </td></tr></tbody></table>

orthotopic liver transplantation in rats

La evolución clínica en el campo del trasplante hepático ha sido ampliamente apoyado por los animales 1,2 modelos. Desde la publicación del primer trasplante ortotópico de hígado de rata en 1979 por Kamada y col. 3, este modelo sigue siendo el estándar de oro a pesar de diversas técnicas propuestas alternativas 4. Sin embargo, su uso más extendido está limitado por su empinada curva de aprendizaje 5. En este trabajo de vídeo, se muestra una revisión simple y fácil de establecer de dos manguito Kamada la técnica. La vena cava suprahepática anastomosis se realiza manualmente con una sutura continua, y la vena porta y la vena cava infrahepática anastomosis se realizan utilizando un sistema de banda rápido enlazador 6. La fabricación del kit de rápida-enlazador se muestra en un papel de video separada.

Watch this Scientific Journal Video about Orthotopic Liver Transplantation in Rats at JoVE.com

Orthotopic Liver Transplantation in Rats

Se presenta una revisión fácil de establecer de la clásica banda de dos técnicas para el trasplante ortotópico de hígado en ratas.

Trasplante ortotópico de hígado en ratas

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

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29.6K vistas.  University of Geneva Hospitals. Se presenta una revisión fácil de establecer de la clásica banda de dos técnicas para el trasplante ortotópico de hígado en ratas.

Video: Orthotopic Liver Transplantation in Rats

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

El desarrollo de bronquiolitis obliterante en un modelo murino de trasplante de pulmón ortotópico

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

Investigación

Medicina

Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

El trasplante ortotópico de intestino delgado en ratas

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

El trasplante ortotópico aórtica en los ratones para el Estudio de la Enfermedad Vascular

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

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 ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

Técnica de Adquisiciones de hígado porcino y trasplante ortotópico usando un Active Porto-Caval derivación

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

Ortotópico de la extremidad posterior de trasplante en el ratón

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

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.

Un modelo de rata de Orthotopic trasplante del hígado usando una técnica de Anastomosis magnética novela para la reconstrucción de la Vena Cava suprahepática

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

Un modelo preclínico porcino de trasplante ortotópico de corazón

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

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.

Reducción de las complicaciones después de la reconexión arterial en un modelo de rata de trasplante hepático ortotópico

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. However, lung transplantation is limited by a shortage of available donor organs. As such, there is high waitlist mortality. Ex situ lung perfusion (ESLP) has increased donor lung utilization rates in some centers by 15%-20%. ESLP has been applied as a method to assess and recondition marginal donor lungs and has demonstrated acceptable short- and long-term outcomes following transplantation of extended criteria donor (ECD) lungs. Large animal (in vivo) transplantation models are required to validate ongoing in vitro research findings. Anatomic and physiologic differences between humans and pigs pose significant technical and anesthetic challenges. An easily reproducible transplant model would permit the&#160;in vivo&#160;validation of current ESLP strategies and the preclinical evaluation of various interventions designed to improve donor lung function. This protocol describes a porcine model of orthotopic left lung allotransplantation. This includes anesthetic and surgical techniques, a customized surgical checklist, troubleshooting, modifications, and the benefits and limitations of the approach.

This protocol describes a juvenile porcine model of orthotopic left lung allotransplantation designed for use with ESLP research. Focus is made on anesthetic and surgical techniques, as well as critical steps and troubleshooting.

Trasplante ortotópico de pulmón izquierdo en un modelo porcino juvenil para LESP

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. ...