Name: heterotopic cervical heart transplantation in mice
Uploaded: 2015-08-25T14:00:00.000+00:00
Duration: 9 min 48 s
Description: Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

M.-A. Deutsch is supported by Dr. Rusche Forschungsprojekt (2014) Deutsche Stiftung f&#252;r Herzforschung and Deutsche Gesellschaft f&#252;r Thorax- Herz- und Gef&#228;&#223;chirurgie. M. Krane is supported by Deutsche Stiftung f&#252;r Herzforschung (F/37/11), Deutsches Zentrum f&#252;r Herz Kreislauf Forschung (DZHK B 13-050A; DZHK B 14-013SE) and Deutsche Forschungsgemeinschaft (KR3770/7-1; KR3770/9-1)

Todos los experimentos fueron aprobados por el alemán Corazón Centro de Múnich y el Gobierno de Baviera, Alemania (Gz. 55.2-1-54-2532.3-17-13). Los animales fueron alojados en una instalación específica libre de patógenos y recibieron atención humana en el cumplimiento de los &quot;Principios de Laboratorio Animal Care&quot; formuladas por la Sociedad Nacional para la Investigación Médica y de la &quot;Guía para el Cuidado y Uso de Animales de Laboratorio&quot; proporcionada por la Academia Nacional de Ciencias y publicado por los Institutos Nacionales de Salud (NIH Publication no. 86-23, revisado 1985). NOTA: Un conjunto completo de instrumentos de microcirugía estériles, incluyendo dos pinzas curvas y dilatadores de los vasos, es necesario para el procedimiento (Figura 2a). La profilaxis antibiótica perioperatoria no es habitualmente necesaria, pero pueden estar indicados en animales inmunocomprometidos. Los animales fueron sacrificados por inhalación de isoflurano usando terminal. Donantes de Preparación y la cosecha del injerto &lt;ol&gt; <li> Anestesiar al animal mediante intraperitoneal (ip) de inyección de midazolam (5,0 mg / kg), medetomidina (0,5 mg / kg) y fentanilo (0,05 mg / kg de peso corporal). Coloque el ratón en una posición supina en el campo operatorio y confirmar la tolerancia quirúrgica por el reflejo de retirada pedal. </li><li> Desinfectar la piel tres veces con clorhexidina y realizar una incisión abdominal en la línea media. Retraer los intestinos con una gasa húmeda a la izquierda para exponer la vena cava inferior. </li><li> Inyectar 50 UI de heparina diluida en 0,4 ml de solución salina a través de la vena cava inferior para la anticoagulación. Incisión en el vaso 1 min después de heparinización. </li><li> Abra el diafragma y realizar una toracotomía bilateral para exponer el corazón. La pared torácica anterior se refleja en sentido superior y se mantiene en su lugar con una abrazadera. </li><li> Retire el timo y sin rodeos diseccionar el tejido conectivo entre la aorta y el tronco pulmonar. Incisión en la vena cava superior derecha. </li><li> Perforar la aorta ascendente movilizado a nivel de latronco braquiocefálico con una aguja de 30 G montado en una jeringa de 5 ml y lentamente perfundir el corazón retrogradually sin una presión excesiva con 3 ml de 4 ° C solución cardiopléjica fría. Evitar la entrada de aire en la circulación coronaria. Caída de solución salina enfriada con hielo en el injerto para mejorar aún más el paro cardiaco. </li><li> Seccionar la aorta proximal al origen del tronco braquiocefálico. </li><li> Ligar la vena inferior y ambas cavas superior cerca del corazón con 8-0 seda y seccionar los distalmente a las ligaduras. </li><li> Diseccionar libre el tronco pulmonar lo más distante posible. Ligar las arterias pulmonares cerca de la bifurcación con 8-0 seda y seccionar ellos distal. </li><li> Ligar las venas pulmonares todos juntos con una sola ligadura de 8-0 seda y seccionar ellos. </li><li> Quite el corazón del donante y almacenarlo en la solución cardiopléjica helada hasta la implantación. </li></ol> 2. Destinatario Preparación <ol><li> Prepararlos puños para anastomosis cortando el tubo de poliimida con un bisturí No. 11 o una tijera bajo el microscopio. El manguito se compone de un cuerpo cilíndrico y una extensión (1/3 de la circunferencia) para la colocación de la pinza vascular, cada uno de una longitud de 1 mm (figura 2b). </li><li> Anestesiar al animal y de confirmar la tolerancia quirúrgica como se describe anteriormente. Inyectar 0,25 ml de solución salina caliente por vía subcutánea (sc) para la reposición de líquidos. </li><li> Quite el pelo de la región cervical derecha usando una máquina de afeitar eléctrica y coloque el animal en posición supina sobre una almohadilla caliente. Asegure las piernas con hebras de cinta evitando estiramiento excesivo de las extremidades anteriores, ya que puede comprometer la respiración. Cubra los ojos utilizando pomada oftálmica para evitar la sequedad. Desinfectar el sitio quirúrgico tres veces con clorhexidina. </li><li> Hacer una incisión cutánea transversal desde el ángulo mandibular derecho a la muesca yugular. </li><li> Sin rodeos movilizar a la extvena yugular ernal. Cauterizar ramas laterales con unas pinzas bipolares y seccionar ellos. </li><li> Divida la vena yugular externa entre ligaduras en el nivel de la confluencia con la vena de la glándula submandibular. </li><li> Tire de la vena yugular externa a través del manguito y ocluir el vaso proximal mediante la colocación de una pinza vascular en la extensión del manguito. Retire la abrazadera y el riego de la luz del vaso con solución salina heparinizada 01:10. </li><li> Evert la pared del vaso sobre el manguito y fijarlo con una circunferencia 8-0 ligadura de seda preestablecido. </li><li> Retire la parte superficial del músculo esternocleidomastoideo. </li><li> Movilizar a la arteria carótida común derecha y seccionar el buque entre ligaduras por debajo de la bifurcación de la carótida. </li><li> Tire de la arteria carótida a través del manguito y ocluir el vaso proximal mediante la colocación de una pinza Yasargil en la extensión del manguito. Retire la abrazadera y el riego de la luz del vaso con solución salina heparinizada 01:10. </li><li> Suavemente dilate la luz arterial con un dilatador buque, Evert la pared del vaso sobre el manguito y arreglarlo con un circunferencial 8-0 ligadura de seda preestablecido. </li><li> Para ganar espacio suficiente para la implantación del injerto, quitar el lóbulo derecho de la glándula submandibular. </li><li> Mantenga el campo quirúrgico húmedo hasta la implantación. </li></ol> 3. Implantación del injerto <ol><li> Coloque el injerto en la región cervical receptor boca abajo con la aorta orientado medialmente y la arteria pulmonar lateralmente. </li><li> Tire de la aorta del injerto sobre el brazalete con la arteria carótida y asegurar la anastomosis con una ligadura circunferencial 8-0 seda. </li><li> Incisión en el tronco pulmonar en su cara anterior y tire de ella sobre el brazalete con la vena yugular externa. Una vez más asegurar la anastomosis con una ligadura circunferencial de 8-0 seda. </li><li> Retire la abrazadera en la vena yugular externa, siguiendo por el desamarre de la arteria carótida. Los corazones se llena inmediatamente con sangre. Para mejorar reperfusion caer 37 ° C una solución salina caliente en el injerto. Normalmente, el corazón se desarrolla el ritmo sinusal en un minuto. Retire la extensión del manguito en el sitio venosa. </li><li> Asegurar la posición correcta del injerto, anastomosis deben estar libre de tensiones y buques sin torsión. No vasos cauterizados de la piel ya que esto puede afectar la cicatrización de heridas. El sangrado generalmente se detiene espontáneamente. Cierre la piel con una sutura continua única de 8-0. </li></ol> 4. Cuidado posoperatorio <ol><li> Inyectar 0,25 ml cálida sc solución salina para la reposición de líquidos. </li><li> Para antagonizar narcóticos inyectan naloxona (1,2 mg / kg), flumazenil (0,5 mg / kg) y atipamezol (2,5 mg / kg de peso corporal) sc Izquierda del animal en la almohadilla caliente hasta que se haya recuperado el conocimiento suficiente para mantener decúbito esternal. Observar al animal durante al menos una hora adicional para registrar cualquier comportamiento anormal. No devuelva un animal que se ha sometido a una cirugía para la compañía de otros animales hasta que esté completamente Recoverojo. </li><li> Proporcionar analgesia preventiva con buprenorfina (0,05 mg / kg sc tres veces por día) o una analgesia alternativa de acuerdo con un protocolo institucional local para una duración mínima de 72 horas después del procedimiento. </li><li> El cirujano y un veterinario deben ver a los animales con regularidad. En caso de pérdida de peso&gt; 15%, la apatía, la hinchazón o animales de infección lado quirúrgicos son sacrificados mediante inhalación de isoflurano terminal. </li></ol>

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K., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+kidney+transplantation+in+mice:+technique+using+cuff+for+renal+vein+anastomosis.">Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis.</a> PloS one. 8, e77278 (2013).</li><li>Maglione, 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=A+novel+technique+for+heterotopic+vascularized+pancreas+transplantation+in+mice+to+assess+ischemia+reperfusion+injury+and+graft+pancreatitis.">A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis.</a> Surgery. 141, 682-689 (2007).</li><li>Sucher, R., 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=Orthotopic+hind-limb+transplantation+in+rats.">Orthotopic hind-limb transplantation in rats.</a> J Vis Exp : JoVE. , (2010).</li><li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp : JoVE. , (2012).</li><li>Okazaki, 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=A+mouse+model+of+orthotopic+vascularized+aerated+lung+transplantation.">A mouse model of orthotopic vascularized aerated lung transplantation.</a> Am J Transplant. 7, 1672-1679 (2007).</li><li>Suzuki, H., Fan, L., Wilkes, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+obliterative+bronchiolitis+in+a+murine+model+of+orthotopic+lung+transplantation.">Development of obliterative bronchiolitis in a murine model of orthotopic lung transplantation.</a> J Vis Exp : JoVE. , (2012).</li><li>Zou, 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=Mouse+model+of+venous+bypass+graft+arteriosclerosis.">Mouse model of venous bypass graft arteriosclerosis.</a> Am J Pathol. 153, 1301-1310 (1998).</li><li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant.">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Exp Clin Transplant. 8, 245-249 (2010).</li><li>Niimi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+technique+for+heterotopic+cardiac+transplantation+in+mice:+experience+of+3000+operations+by+one+surgeon.">The technique for heterotopic cardiac transplantation in mice: experience of 3000 operations by one surgeon.</a> J Heart Lung Transplant. 20, 1123-1128 (2001).</li><li>Arras, M., Autenried, P., Rettich, A., Spaeni, D., Rulicke, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+intraperitoneal+injection+anesthesia+in+mice:+drugs,+dosages,+adverse+effects,+and+anesthesia+depth.">Optimization of intraperitoneal injection anesthesia in mice: drugs, dosages, adverse effects, and anesthesia depth.</a> Comp Med. 51, 443-456 (2001).</li></ol>

The authors have nothing to disclose.

Hemos completado más de 200 trasplantes cardíacos con una tasa de éxito superior al 90% en varios parámetros experimentales. Con la técnica descrita aquí, el procedimiento completo se puede hacer dentro de los 60 min por un cirujano experimentado. Mediante la preparación de los vasos cervicales antes de la obtención de órganos, el tiempo total de isquemia del injerto puede ser limitado a 20 min. Tiempo para la implantación (tiempo de isquemia caliente) es significativamente más corto usando la técnica del ma...

El modelo de trasplante de corazón se utiliza con frecuencia para investigar I / R-lesión, rechazo de aloinjerto, vasculopatía post-trasplante y la eficiencia de los agentes inmunomoduladores. Los modelos de ratón se combinan muchas ventajas como el inmunológico conocido y antecedentes genéticos, la disponibilidad de muchas cepas puras y transgénicos y los costos experimentales relativamente bajos. En 1973, una técnica de trasplante de corazón abdominal en ratones fue descrita por primera vez por Corry et al 1. En este modelo injertos se revascularizados antes de fin de a lado anastomosis de la aorta ascendente a la aorta abdominal del receptor y de la principal arteria pulmonar a la vena cava inferior. En 1991, una técnica de sutura para el trasplante de corazón cervical en ratones fue descrito por Chen et al 2. En este modelo una perfusión coronaria retrógrada se establece por la anastomosis de la arteria carótida del destinatario a la aorta ascendente del grpopa. Los drenajes de sangre venosa a través del seno coronario en la aurícula derecha, el ventrículo derecho y se expulsa a través de la arteria pulmonar en el receptor vena yugular externa (Figura 1). En comparación con la implantación del injerto abdominal el modelo de cuello uterino es menos invasiva y la localización superficial del injerto permite un acceso muy fácil para los exámenes de seguimiento adicionales (por ejemplo, la palpación, la ecocardiografía, la microscopía de fluorescencia intravital) 3,4. A pesar de las mejoras técnicas 5,6, un uso más generalizado de este modelo ha sido limitado debido a dificultades técnicas para realizar las anastomosis suturadas de vasos pequeños con una alta incidencia de fugas de anastomosis y la trombosis. En 1991, Matsuura et al. introdujo la técnica de manguito en la que la pared del vaso es evertido sobre un cilindro sintético, que facilita en gran medida el modelo de trasplante de corazón 7 cervical. La técnica fue further adoptado para varios modelos experimentales de trasplante renal incluyendo 8, 9 de páncreas, la integridad física 10, 11,12 hepática, pulmonar 13,14 y el trasplante vascular 15. En comparación con la técnica de sutura el tiempo requerido para las anastomosis y, por lo tanto, el tiempo de isquemia caliente es más corta y hay complicaciones vasculares significativamente menos usando la técnica del manguito 16. Además, permite a los cirujanos con poca formación en microcirugía para realizar la operación con una alta tasa de éxito. Una desventaja de la técnica del manguito es el requisito previo de la ligadura de la arteria carótida común, que puede alterar la perfusión cerebral. En este trabajo de vídeo presentamos una técnica de manguito mejorado y simplificado para el trasplante de corazón cervical en ratones.

<table><tbody><tr><td>Forceps JFC-7.2</td><td>S&amp;amp;T</td><td>P-00036</td><td>Curved tip 0.20 mm</td></tr><tr><td>Vessel dilatator D-5a</td><td>S&amp;amp;T</td><td>S-00124</td><td></td></tr><tr><td>Adventita scissor</td><td>S&amp;amp;T</td><td>S-00102</td><td></td></tr><tr><td>Vascular clamp B-1V</td><td>S&amp;amp;T</td><td>S-00396</td><td></td></tr><tr><td>Yasargil Clip</td><td>Peter Lazic</td><td>65,097</td><td>3.5 mm lenght</td></tr><tr><td>Bipolar forceps</td><td>Micromed</td><td>148-100-011</td><td>tip 0.25 mm</td></tr><tr><td>Polyimide tubes</td><td>River Tech Medical</td><td></td><td>19-24 gauge</td></tr><tr><td>8.0 silk ligatures</td><td>Catgut</td><td></td><td></td></tr><tr><td>Custodiol HTK solution</td><td>Essential Pharmaceuticals</td><td></td><td></td></tr></tbody></table>

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.

Nuestro equipo podría realizar con éxito la técnica descrita después de aproximadamente 20 cirugías de formación. La parte más difícil es eversión la pared vascular de la arteria carótida sobre el brazalete, ya que fácilmente voltea hacia atrás y múltiples intentos puede conducir a la rotura de la pared. Después del período de entrenamiento, el tiempo quirúrgico total media fue de 60 ± 8 min. Se requieren alrededor de 20 minutos para la cosecha de donantes, 25 min para la preparación de los vasos cervi...

Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

Heterotopic Cervical Heart Transplantation in Mice

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 ...

heterotopic-cervical-heart-transplantation-in-mice

Research

JoVE Journal

Medicine

11.1K vistas.  German Heart Center Munich, Technische Universität München. 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.

Video: Heterotopic Cervical Heart Transplantation in Mice

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

El trasplante cardíaco heterotópico en ratones

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

Investigación

Biología

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

Murino Cervical Trasplante Cardiaco Modelo Usando una técnica de Cuff Modificado

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

Medicina

A Modified Method for Heterotopic Mouse Heart Transplantion

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents available. A difficulty is presented due to the small size of the animal and the considerable technical challenges of the microsurgery involved in heart transplantation. In particular, a high rate of technical failure early after transplantation may result from recipient death and post-operative complications such as hind limb paralysis or a non-beating heart. Here, the complete technique for heterotopic mouse heart transplantation is demonstrated, involving harvesting the donor heart and its subsequent implantation into a recipient mouse. The donor heart is harvested immediately following in situ perfusion with cold heparinized saline and transection of the ascending aorta and pulmonary artery. The recipient operation involves preparation of the abdominal aorta and inferior vena cava (IVC), followed by end-to-side anastomosis of the donor aorta with the recipient aorta using a single running 10-0 microsuture and a similar anastomosis of the donor pulmonary artery with the recipient IVC. Following the operation the animal is injected with 0.6 ml normal saline subcutaneously and allowed to recover on a 37&#160;&#176;C heating pad. The results from 227 mouse heart transplants are summarized with a success rate at 48 hr of 86.8%. Of the 13.2% failures within 48 hr, 5 (2.2%) experienced hind limb paralysis, 10 (4.4%) had a non-beating heart due to graft ischemic injury and/or thrombosis, while 15 (6.6%) died within 48 hr.

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

Un método modificado para heterotópico Ratón Corazón TRASPLANTE

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

Murino heterotópico Corazón Trasplante Técnica

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

Un microquirúrgica Modelo Novela de heterotópico, en bloque, pared torácica, el timo y el trasplante cardiaco en ratones

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

An Immunological Model for Heterotopic Heart and Cardiac Muscle Cell Transplantation in Rats

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications have been reported since the first description in 1964. After 30 years of performing heterotopic heart transplantation in rats, we have developed a simplified surgical approach, which can be easily taught and performed without further surgical training or background.
After dissection of the ascending aorta and the pulmonary artery and ligation of superior and inferior caval and pulmonary veins, the donor heart is harvested and subsequently perfused with ice-cold saline solution supplemented with heparin. After clamping and incising the recipient abdominal vessels, the donor ascending aorta and pulmonary artery are anastomosed to the recipient abdominal aorta and inferior vena cava, respectively, using continuous running sutures.
Depending on different donor-recipient combinations, this model allows analyses of either acute or chronic rejection of allografts. The immunological significance of this model is further enhanced by a novel approach of in-ear injection of vital cardiac muscle cells and subsequent analysis of draining cervical lymphatic tissue.

We describe a model of heterotopic abdominal heart transplantation in rats, implying modifications of current strategies, which lead to a simplified surgical approach. Additionally, we describe a novel rejection model by in-ear injection of vital cardiac muscle cells, allowing further transplant immunological analyses in rats.

Un modelo inmunológico para el trasplante heterotópico de células musculares cardíacas y cardíacas en ratas

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications ...

Inmunología e Infección

Optimization of the Cuff Technique for Murine Heart Transplantation

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

Optimización de la Técnica del Manguito para el Trasplante cardíaco murino

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to ...

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 ...

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

Trasplante cardíaco cervical heterotópico de ratón utilizando manguitos vasculares

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

Reconstrucción Del Circuito Sanguíneo En Un Modelo De Trasplante De Corazón De Ratón Abdominal

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...