1. Donante de Operación <ol><li> La rata donante debe mantenerse en ayunas durante 24 horas (acceso libre a una solución de agua / glucosa). </li><li> Para inducir la anestesia por inhalación isoflurano, comience con un 2% en atomizador estándar, y luego reducir a 1% después de realizar una laparotomía. Realice una pizca dedo para comprobar la sedación. </li><li> Afeitar el abdomen y limpiar con preparación de la piel 3 veces (Kodan). A continuación, realizar una incisión media después de la administración subcutánea del analgésico. </li><li> Después de que el injerto se envuelve en una gasa empapada en solución salina, separar las adherencias fisiológicas entre el páncreas y el colon ascendente suavemente con un Q-tip (bajo el microscopio quirúrgico con el aumento de 6x). </li><li> Ligar y dividir los vasos cólicos ileocecal y derecho y medio con 7-0 seda. Después de que el colon ascendente se extiende hacia el lado derecho de la vena mesentérica superior (SMV), los vasos cólicos ileocecal, central y derecha se pueden identificar para la ligadura y división, usando 7-0 lazos. </li><li> Retraer el estómago hacia arriba, de modo que la SMV todo se endereza y expuesta. Utilice una pinza mosquito para la retracción. La pinza se mantiene en masa de plastilina formado en forma según sea necesario. </li><li> Ligar y dividir las venas pancreático-duodenales procedentes de la SMV. Todos los pequeños pancreático-duodenales venas procedentes de la SMV debe identificarse cuidadosamente, se ligó con seda 7-0 y dividida antes de que el tejido pancreático se puede quitar desde el injerto. </li><li> Ligar y dividir el tejido conectivo incluyendo perder todos los vasos linfáticos entre el SMV y la aorta abdominal. Con la prótesis todavía en el lado derecho del abdomen, el perder tejido conectivo incluyendo todos los vasos linfáticos entre el SMV y la aorta abdominal es accesible. Este tejido conectivo deben ser ligados utilizando seda 7-0 y se divide para evitar linforrea postoperatoria del injerto intestinal. </li><li> Ligar y dividir la arteria renal derecha. Después de que el tejido conectivo se divide, la arteria renal derecha se convierte en accessible y se liga y se divide usando seda 7-0. </li><li> Sistémicamente heparinise la rata usando 200 unidades de heparina iv a través de la vena del pene. </li><li> Ligar las arterias marginales, y dividir el intestino delgado en la unión duodeno-yeyunal y en el íleon terminal. </li><li> La aorta se liga proximal al origen de la SMA. La vena porta se secciona en la confluencia con la vena esplénica. A continuación, el injerto se cosecha con su pedículo vascular que consiste de la SMA con un segmento de la aorta. </li></ol> 2. Procedimiento Backtable <ol><li> Perfundir el injerto usando 3 ml de solución de la Universidad de Wisconsin (UW) a 4 ° C a través del conducto aórtico e irrigar el lumen intestinal desde el extremo yeyunal con 30 ml de Uro-Nebacetin solución de N a 4 ° C (lumen de irrigación es obligatorio) . </li></ol> Inmediatamente después de extraer el injerto, el conducto aórtico se utiliza para la perfusión con 3 ml de solución fría de UW. Para ello, un 20 G iv catéter enuna jeringa de 10 ml se utiliza. El perfundido se debe observar para fluir libremente desde la vena portal dividido. Para la irrigación intestinal con Nebacetin, una jeringa de 50 ml se utiliza. <ol start="2"><li> Almacenar el injerto en solución UW a 4 º C durante la preparación del receptor. </li></ol> 3. Receptor Operación <ol><li> La rata receptora debe mantenerse en ayunas durante 24 horas (acceso libre a una solución de agua / glucosa). </li><li> Para inducir la anestesia por inhalación isoflurano, comience con un 2% en atomizador estándar, y luego reducir a 1% después de realizar una laparotomía. Realice una pizca dedo para comprobar la sedación. </li><li> Afeitar el abdomen y limpiar con preparación de la piel 3 veces (Kodan). A continuación, realizar una incisión media después de la administración subcutánea de Carprofen 5 mg / kg sc de analgesia. </li><li> Envuelva el recipiente en condiciones normales del intestino gasa empapada en solución salina y se coloca en el pecho del receptor. </li><li> Abra el retroperitoneo sin rodeos con Q-tips, y exponer el abdominal aorta y vena cava inferior justo por debajo de los vasos renales hasta el nivel de los vasos ilíacos, si es necesario también utilizar microtijeras. Se liga el tribuaries pequeñas espinal lumbar y de la aorta y la vena cava utilizando 7-0 seda para evitar la pérdida de sangre. (Para ello, cambie el zoom de 16x microscopio quirúrgico.) </li><li> Pinzamiento de la aorta y la vena cava inferior por debajo de la de los vasos renales proximales izquierdas y por encima de la bifurcación ilíaca distal utilizando clips de microvasos. Sólo una de las pinzas se utiliza proximalmente como distalmente así como para sujetar los dos buques simultáneamente. Una incisión ambos buques anteriormente utilizando un microknife y lavar la sangre restante. </li><li> Crear una anastomosis de extremo a lado entre el segmento aórtico injerto y la aorta infrarrenal del destinatario mediante una continua 10-0 sutura Prolene .. Inicialmente, el injerto se coloca en el lado derecho del abdomen (la cabeza de la rata colocada en las 12 horas) para realizar las puntadas de la pared posterior de la anastomosis arterial y la vinculación de la sutura estancia inferior. Then, el injerto se gira hacia el lado izquierdo del abdomen (la cabeza de la rata colocada en las 9 horas) para exponer y suturar la pared frontal de la anastomosis. </li><li> Una anastomosis de extremo a lado entre la vena porta y la vena cava inferior del injerto del destinatario se realiza igualmente mediante la ejecución de suturas utilizando 10-0 Prolene. Con la rata fijas situadas lateralmente (cabeza en posición de las 9), el injerto se coloca en el lado izquierdo del abdomen y una sutura de estancia inferior se coloca. La anastomosis se inicia con la pared posterior desde el interior del recipiente. Después de la sutura estancia inferior está ligado, las puntadas de la pared principal se puede realizar desde el exterior. </li><li> Retire las abrazaderas distales en primer lugar, seguidos de las abrazaderas superiores. Cualquier hemorragia anastomótica es controlado por la presión directa con un Q-tips. El injerto debe ser revisado por reperfusión igual y rápida. </li><li> Resecar el intestino delgado del receptor entero después de la ligadura de los vasos mesentéricos. Los beneficiarios se someten enterectomía subtotal, preservando 2-3 cm de proyeyuno ximal y 1 cm de íleon distal. </li><li> Restaurar la continuidad intestinal por proximal (yeyuno-yeyunostomía) y distal (ileo-ileostomía) de extremo a extremo utilizando una anastomosis intestinales interrumpido de una capa de sutura con 6-0 Monocryl. Aproximadamente 16 puntos de sutura se necesitan para completar las anastomosis. </li><li> Irrigar la cavidad peritoneal con solución salina normal hasta que esté limpia. Administrar 2 ml de solución salina normal por vía intraperitoneal para la reposición de líquidos. A continuación, cierre el abdomen con sutura continua con 3-0 Vicryl para la capa muscular más una sutura de la piel continua. </li><li> En el postoperatorio, las ratas se debe mantener en ayunas (con acceso al agua y una solución de glucosa) por otro de 24 horas luego reanudaron el estándar rata chow y agua ad libitum. Analgesia con carprofeno se debe administrar durante 3 días (ver dosis más adelante). </li></ol>

<ol>
	<li>Abu-Elmagd, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Five+Hundred+Intestinal+and+Multivisceral+Transplantations+at+a+Single+Center:+Major+Advances+With+New+Challenges.">Five Hundred Intestinal and Multivisceral Transplantations at a Single Center: Major Advances With New Challenges.</a> Ann. Surg. 250 (4), 567-56 (2009).</li><li>Berney, T., 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+versus+systemic+drainage+of+small+bowel+allografts:+comparative+assessment+of+survival,+function,+rejection,+and+bacterial+translocation.">Portal versus systemic drainage of small bowel allografts: comparative assessment of survival, function, rejection, and bacterial translocation.</a> J. Am. Coll. Surg. 195 (6), 804 (2002).</li><li>Fishbein, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+transplantation.">Intestinal transplantation.</a> N. Engl. J. Med. 361 (10), 998 (2009).</li><li>Hernandez, F., 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=Is+portal+venous+outflow+better+than+systemic+venous+outflow+in+small+bowel+transplantation?+Experimental+study+in+syngeneic+rats.">Is portal venous outflow better than systemic venous outflow in small bowel transplantation? Experimental study in syngeneic rats.</a> J. Pediatr. Surg. 40 (2), 336 (2005).</li><li>Monchik, G. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+small+bowel+in+the+rat:+technical+and+immunological+considerations.">Transplantation of small bowel in the rat: technical and immunological considerations.</a> Surgery. 70 (5), 693 (1971).</li><li>Murase, N., 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=Graft-versus-host+disease+after+brown+Norway-to-Lewis+and+Lewis-to-Brown+Norway+rat+intestinal+transplantation+under+FK506.">Graft-versus-host disease after brown Norway-to-Lewis and Lewis-to-Brown Norway rat intestinal transplantation under FK506.</a> Transplantation. 55 (1), 1 (1993).</li><li>Murase, N., 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=Immunomodulation+of+intestinal+transplant+with+allograft+irradiation+and+simultaneous+donor+bone+marrow+infusion.">Immunomodulation of intestinal transplant with allograft irradiation and simultaneous donor bone marrow infusion.</a> Transplant Proc. 31 (1-2), 565 (1999).</li><li>Pech, T., 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=Perioperative+infliximab+application+ameliorates+acute+rejection+associated+inflammation+after+intestinal+transplantation.">Perioperative infliximab application ameliorates acute rejection associated inflammation after intestinal transplantation.</a> Am. J. Transplant. 10 (11), 2431 (2010).</li><li>Schaefer, N., 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=Resident+macrophages+are+involved+in+intestinal+transplantation-associated+inflammation+and+motoric+dysfunction+of+the+graft+muscularis.">Resident macrophages are involved in intestinal transplantation-associated inflammation and motoric dysfunction of the graft muscularis.</a> Am J Transplant. 7 (5), 1062 (2007).</li><li>Schaefer, N., 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=Mechanism+and+impact+of+organ+harvesting+and+ischemia-reperfusion+injury+within+the+graft+muscularis+in+rat+small+bowel+transplantation.+Transplant.">Mechanism and impact of organ harvesting and ischemia-reperfusion injury within the graft muscularis in rat small bowel transplantation. Transplant.</a> Proc. 38 (6), 1821 (2006).</li><li>Schaefer, N., 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=Acute+rejection+and+the+muscularis+propria+after+intestinal+transplantation:+the+alloresponse,+inflammation,+and+smooth+muscle+function.">Acute rejection and the muscularis propria after intestinal transplantation: the alloresponse, inflammation, and smooth muscle function.</a> Transplantation. 85 (10), 1465 (2008).</li><li>Xue, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+experience+of+refined+3-cuff+technique+for+orthotopic+small-bowel+transplantation+in+rat:+a+report+of+270+cases.">Surgical experience of refined 3-cuff technique for orthotopic small-bowel transplantation in rat: a report of 270 cases.</a> Am. J. Surg. 198 (1), 110 (2009).</li><li>Zhang, X. 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=Simplified+techniques+in+rat+heterotopic+small+bowel+transplantation.">Simplified techniques in rat heterotopic small bowel transplantation.</a> Transplant. Proc. 38 (6), 1840 (2006).</li><li>Zhao, Y., 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=The+establishment+of+a+new+en+bloc+combined+liver-small+bowel+transplantation+model+in+rats.+Transplant.">The establishment of a new en bloc combined liver-small bowel transplantation model in rats. Transplant.</a> Proc. 39 (1), 278 (2007).</li></ol>

No hay conflictos de interés declarado.

Mientras que los modelos de trasplante intestinal en ratas se han descrito ya en el 1970ies 5 la mayoría de los modelos recientemente empleadas implican trasplante heterotópico intestinal utilizando diferentes técnicas 13. Si bien los procedimientos heterotópico en general, tienen la ventaja de las técnicas de microcirugía más y más fácil accesibilidad del injerto para la evaluación, el trasplante heterotópico intestinal tiene la gran desventaja de no tener en cuenta las múltiples inter...

<table border="1"><tbody><tr><td> Nombre del reactivo </td><td> Empresa </td><td> Número de catálogo </td><td> Comentarios (opcional) </td></tr><tr><td> Universidad de Wisconsin (UW) solución (ViaSpan) </td><td> Bristol-Myers Squibb </td><td></td><td></td></tr><tr><td> Uro-N Nebacetin solución </td><td> Nycomed </td><td> 6967855 </td><td></td></tr><tr><td> Ampicilina </td><td> Ratiopharm </td><td></td><td></td></tr><tr><td> Carprofeno (Rimadyl) </td><td> Pfizer </td><td></td><td></td></tr><tr><td> 10-0 sutura Prolene unresorbable </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Monocryl 6-0 sutura reabsorbible </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Vicryl 3-0 sutura reabsorbible </td><td> Ethicon </td><td></td><td></td></tr><tr><td> iv catéter G 20 mm 1.1x33 </td><td> Braun </td><td></td><td></td></tr><tr><td> El catéter iv G 22 0.9x25 mm </td><td> Braun </td><td></td><td></td></tr><tr><td> Kodan Skin Prep </td><td> Schülke </td><td></td><td></td></tr><tr><td> Infusión NaCl 0,9% de solución </td><td> Braun </td><td></td><td></td></tr><tr><td> Pinzas curvas pequeña </td><td> FineScienceTools </td><td> 11009-13 </td><td></td></tr><tr><td> Micro pinzas curvas </td><td> AESCULAP </td><td> BD 333 </td><td></td></tr><tr><td> Micro pinzas curvas </td><td> AESCULAP </td><td> FD281R </td><td></td></tr><tr><td> Micro Pinza recta 1 </td><td> WPI </td><td> 5 </td><td></td></tr><tr><td> Micro Pinza recta 2 </td><td> WPI </td><td> 2 </td><td></td></tr><tr><td> Soporte de la aguja Micro </td><td> WPI </td> &lt;td&gt; 14081 </td><td></td></tr><tr><td> Micro tijeras </td><td> FineScienceTools </td><td> 15006-09 </td><td></td></tr><tr><td> Micro bisturí </td><td> MANI </td><td> Oftálmica cuchillo </td><td></td></tr><tr><td> Micro pinzas </td><td> AESCULAP </td><td> FB329R </td><td></td></tr></tbody></table>

orthotopic small bowel transplantation in rats

Trasplante de intestino delgado se ha convertido en una opción aceptada clínica de los pacientes con síndrome de intestino corto y la falta de nutrición parenteral (fallo intestinal irreversible). En los centros especializados mejorado la gestión operativa y estrategias han llevado a excelente a corto y medio plazo paciente y la supervivencia del injerto al tiempo que proporciona alta calidad de vida 1,3. A diferencia del trasplante más común de otros órganos sólidos (corazón, es decir, hígado) muchos mecanismos subyacentes de la función del injerto y las alteraciones inmunológicas inducidas por el trasplante intestinal no son del todo conocidos 6,7. Los episodios de rechazo agudo, sepsis e insuficiencia crónica del injerto son los principales obstáculos aún menos favorable que contribuyen al resultado a largo plazo y dificultando un empleo más generalizado del procedimiento a pesar de un número creciente de pacientes con nutrición parenteral domiciliaria que potencialmente se beneficiarían de un trasplante. El intestino delgado contieneun gran número de leucocitos pasajeros comúnmente como parte del sistema linfoide asociado a intestino (GALT) esta siendo parte de la razón de la alta immunogenity del injerto intestinal. La presencia y la proximidad de muchos comensales y patógenos en el intestino explica la gravedad de los episodios de sepsis una vez integridad de la mucosa del injerto se ve comprometida (por ejemplo, rechazo). Para avanzar en el campo de datos intestinales y trasplante multiorgánico más generados a partir de modelos animales fiables y factibles que se necesita. El modelo que aquí combina la fiabilidad y viabilidad una vez establecido de una manera estandarizada y pueden proporcionar información valiosa en los subyacentes complejos mecanismos moleculares, celulares y funcionales que se activan mediante el trasplante intestinal. Hemos utilizado con éxito y perfeccionado el procedimiento descrito a lo largo de más de 5 años en nuestro laboratorio 8-11. El JoVE basado en video formato es especialmente útil para demostrar el procedimiento complejoDuré y evitar los errores iniciales de los grupos de planificación para establecer un modelo de roedor ortotópico investigando el trasplante intestinal.

Curso postoperatorio normal Los animales trasplantados deben recuperarse rápidamente del procedimiento bajo una lámpara de calor durante aproximadamente 1 hora. La hipotermia es una causa importante de mortalidad postoperatoria y debe evitarse cuidadosamente intra-y postoperatorio. Pérdidas intraoperatorias de fluido debe ser sustituido por inyección subcutánea de 2,5 ml de solución salina normal más 2,5 ml de glucosa al 5% cada 8 horas durante las primeras 24 h. Las ratas también deben...

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

Orthotopic Small Bowel Transplantation in Rats

Trasplante de intestino delgado se ha convertido en una opción terapéutica en los pacientes con fallo intestinal irreversible. Nuestro modelo experimental de trasplante ortotópico del intestino delgado en ratas sirve como una herramienta fiable para abordar inmunológica subyacente y los procesos inflamatorios que complicar el trasplante intestinal.

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-small-bowel-transplantation-in-rats

Research

JoVE Journal

Medicine

14.1K vistas.  University of Bonn, Germany. Trasplante de intestino delgado se ha convertido en una opción terapéutica en los pacientes con fallo intestinal irreversible. Nuestro modelo experimental de trasplante ortotópico del intestino delgado en ratas sirve como una herramienta fiable para abordar inmunológica subyacente y los procesos inflamatorios que complicar el trasplante intestinal.

Video: Orthotopic Small Bowel 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 Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

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 ...

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. ...

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 ...

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis. In recent years, oncolytic virotherapies have proven themselves to be potentially powerful tools to fight malignancy. Due to the unique dual blood supply in the liver, it is possible to apply therapies locally to orthotopic liver tumors, which are predominantly fed by arterial blood flow. We have previously demonstrated that hepatic arterial delivery of oncolytic viruses results in safe and efficient transduction efficiency of multifocal HCC lesions, resulting in significant prolongation of survival in immune competent rats. This procedure closely mimics the application of transarterial embolization in patients, which is the standard palliative care provided to many HCC patients. The ability to administer tumor therapies through the hepatic artery in rats allows for a highly sophisticated preclinical model for evaluating novel viral vectors under development. Here we describe the detailed protocol for microdissection of the hepatic artery for infusion of oncolytic virus vectors to treat orthotopic HCC.

Oncolytic virotherapies are under development as novel therapeutics for the treatment of hepatocellular carcinoma (HCC). Here we describe a method for locoregional therapy of HCC via hepatic arterial administration of oncolytic virus.

Administración transarterial de virus oncolíticos para locorregional Terapia de HCC ortotópico en ratas

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis.  In recent years, oncolytic virotherapies have proven ...

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 ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

Trasplante de riñón de rata ortotópica: un enfoque quirúrgico novedoso y simplificado

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

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 ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

Autoaplano de riñón ortotópico en un modelo porcino utilizando 24 horas de preservación de órganos y telemetría continua

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...

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. ...