The authors declare that they have no competing financial interests.

Los animales se alojan en un centro sin barreras patógeno. Todos los animales reciben atención humana en el cumplimiento de los &quot;Principios de Cuidado de Animales de Laboratorio&quot; formuladas por la Sociedad Nacional para la Investigación Médica &quot;y la&quot; Guía para el Cuidado y Uso de Animales de Laboratorio &quot;, elaborado por la Academia Nacional de Ciencias y publicados por la National Institutos de Salud (NIH Publication no. 86-23, revisado 1985). Todos los experimentos son aprobados por el Ministerio de Educación, Ciencia y Cultura de Austria. 1. destinatario Preparación <ol><li> Anestesiar al animal receptor con una inyección im de xilazina (5 mg / kg de peso corporal) y ketamina (100 mg / kg de peso corporal). </li><li> Quite todo el pelo de la región cervical lateral del animal y fregar el campo operatorio tres veces usando chlorhexdine. </li><li> Coloque el ratón en una posición supina en el campo operatorio. </li><li> Siguiente hacer una incisión en la piel del inc yugularisión de la mandíbula inferior derecha. </li><li> Posteriormente, sin rodeos movilizar la vena yugular externa derecha (VYE) y dividir las ramas entre ligaduras. </li><li> Luego divida la VYE entre ligaduras asegurándose de dejar suficiente longitud para eversión del muñón proximal sobre el cuerpo del manguito. </li><li> A continuación, retire el lóbulo derecho de la glándula submandibular. </li><li> Para la anastomosis venosa pasar el extremo proximal de la VYE a través del manguito de polietileno y fijarlo en el mango del manguito con una abrazadera micro. </li><li> Retire la ligadura en el extremo de la embarcación, eversión de la luz sobre el manguito y fijarla con un 8-0 lazo de seda (Figura 1a). </li><li> Posteriormente seccionar el músculo esternocleidomastoideo derecha con cauterio bipolar para obtener acceso a la arteria carótida común. </li><li> Siguiente movilizar la arteria carótida común y cortar el vaso entre las ligaduras. </li><li> Siguiente movilizar la arteria carótida común y cortar el vaso entre las ligaduras. </li><li> Pase los disTal extremo del recipiente a través del manguito y fijarlo con la abrazadera arteria. </li><li> Retire la ligadura en el extremo del recipiente y distender el lumen usando dilatadores vasculares. A continuación, en analogía con el VYE, eversión de la arteria en el manguito y fijarla con un lazo de seda 8-0. </li></ol> Contratación 2. Corazón <ol><li> Anestesiar el ratón donante con una inyección im de xilazina (5 mg / kg de peso corporal) y ketamina (100 mg / kg de peso corporal). A continuación, retire todo el pelo de la región abdominal del animal. Frote el operativo de campo tres veces usando chlorhexdine. Siguiente lugar el ratón en una posición supina en el campo quirúrgico como se describió previamente. </li><li> Después de una incisión abdominal en la línea media, retraiga las vísceras con Q-tips a la izquierda con el fin de exponer la vena cava inferior (VCI). </li><li> Inyectar 400 l de una mezcla 1: 4 heparina solución de sodio en el IVC para la heparinización. </li><li> Lleve a cabo una toracotomía y doblar la pared anterior del tórax sobre craneal para ganar access al corazón. </li><li> A continuación, retire el timo y hacer venteo incisiones en la vena cava superior izquierda y la derecha. </li><li> Posteriormente, perfundir el corazón de una manera retrógrada con solución de 4 ° Celsius HTK por canulación del arco aórtico con una G jeringa 27 en el nivel de la rama braquiocefálico. </li><li> Ligar la vena cava inferior y superior con 8-0 seda y dividirlos distal a las ligaduras. </li><li> Siguiente dividir el arco aórtico en el nivel de la canulación anterior y diseccionar libre del tronco pulmonar y se divide distalmente en la medida de lo posible. </li><li> Después de eso, atar las venas pulmonares con una ligadura en bloque y dividirlos distal a las ligaduras. </li><li> Por último, quite el corazón de la zona donante y almacenarlo en 4 ° centígrados solución HTK (Figura 1b). </li></ol> 3. Implantación <ol><li> Coloque el injerto de corazón en la región del cuello destinatario en una posición al revés (Figura 1c </strong&gt;). </li><li> Siguiente tirar del tronco pulmonar del corazón sobre la construcción del manguito vena del receptor y fijarlo con un lazo de seda (Figura 1d). </li><li> Realizar la anastomosis entre la aorta del injerto y la arteria evertido del animal receptor de la misma manera (Figura 1e). </li><li> Posteriormente, retire la pinza venosa primero, seguido por la pinza arterial. El corazón está reperfunde y comienza a latir dentro de 1-2 min. Durante la reperfusión humedecer el corazón con agua tibia (35 ° Celsius) solución salina. </li><li> Por último cerrar la herida quirúrgica con 6-0 suturas continuas (Figura 1e). </li></ol> 4. Cuidado posoperatorio, punto final <ol><li> Dé hasta 0,3 ml ip salina normal después de la operación para la reposición de líquidos. </li><li> Colocar el animal bajo una lámpara de calor hasta que despierta de la anestesia. </li><li> Una vez despierto, devolver el animal a la instalación de la vivienda donde reciben alimentos y agua ad libitum. </li><li> Para midolor de quitar importancia durante los primeros 7 días del postoperatorio administrar carprofeno (4 mg / kg por vía subcutánea cada 12 horas (sc). Además, dará buprenorfina (0,1 mg / kg) después de la operación cada 12 horas durante 5 días. </li><li> Obtener el peso (g) de cada destinatario de los animales semanalmente para evaluar la ingesta nutricional apropiada. Si hay evidencia de pérdida de peso más de un 10-15% en comparación con el peso en la cirugía fecha, la apatía, la paralizante o una espalda muy doblada, sacrificar al animal mediante la inhalación de isoflurano terminal antes de llegar al punto final clínico. También sacrificar al animal después de que el corazón ha sido rechazada. </li></ol>

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+mechanisms+of+cardiac+allograft+vasculopathy+an+update.">Immune mechanisms of cardiac allograft vasculopathy an update.</a> Transpl Immunol. 1, 237-249 (1993).</li><li>Libby, P., Pober, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+rejection.">Chronic rejection.</a> Immunity. 14, 387-397 (2001).</li><li>Kouwenhoven, E. A., de Bruin, R. W., Heemann, U. W., Marquet, R. L., Ijzermans, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Late+graft+dysfunction+after+prolonged+cold+ischemia+of+the+donor+kidney+inhibition+by+cyclosporine.">Late graft dysfunction after prolonged cold ischemia of the donor kidney inhibition by cyclosporine.</a> Transplantation. 68, 1004-1010 (1999).</li><li>Kouwenhoven, E. A., Ijzermans, J. N., de Bruin, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiology+and+pathophysiology+of+chronic+transplant+dysfunction.">Etiology and pathophysiology of chronic transplant dysfunction.</a> Transpl Int. 13, 385-401 (2000).</li><li>Chen, Z. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+of+cervical+heterotopic+heart+transplantation+in+mice.">A technique of cervical heterotopic heart transplantation in mice.</a> Transplantation. 52, 1099-1101 (1991).</li><li>Huang, X., Chen, D., Chen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=[A+modified+model+of+cervical+heterotopic+cardiac+transplantation+for+chronic+rejection+research].">[A modified model of cervical heterotopic cardiac transplantation for chronic rejection research].</a> Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 22, 1508-1510 (2008).</li><li>Hasegawa, T., Visovatti, S. H., Hyman, M. C., Hayasaki, T., Pinsky, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+vascularized+murine+cardiac+transplantation+to+study+graft+arteriopathy.">Heterotopic vascularized murine cardiac transplantation to study graft arteriopathy.</a> Nat Protoc. 2, 471-480 (2007).</li><li>Wang, C. 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=Suppression+of+murine+cardiac+allograft+arteriopathy+by+long-term+blockade+of+CD40-CD154+interactions.">Suppression of murine cardiac allograft arteriopathy by long-term blockade of CD40-CD154 interactions.</a> Circulation. 105, 1609-1614 (2002).</li><li>Matsuura, A., Abe, T., Yasuura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+mouse+cervical+heart+transplantation+using+a+cuff+technique.">Simplified mouse cervical heart transplantation using a cuff technique.</a> Transplantation. 51, 896-898 (1991).</li><li>Tomita, 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=Improved+technique+of+heterotopic+cervical+heart+transplantation+in+mice.">Improved technique of heterotopic cervical heart transplantation in mice.</a> Transplantation. 64, 1598-1601 (1997).</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>Corry, R. J., Winn, H. 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=Primarily+vascularized+allografts+of+hearts+in+mice.+The+role+of+H-2D,+H-2K,+and+non-H-2+antigens+in+rejection.">Primarily vascularized allografts of hearts in mice. The role of H-2D, H-2K, and non-H-2 antigens in rejection.</a> Transplantation. 16, 343-350 (1973).</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>Zimmermann, F. A., 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=Techniques+for+orthotopic+liver+transplantation+in+the+rat+and+some+studies+of+the+immunologic+responses+to+fully+allogeneic+liver+grafts.">Techniques for orthotopic liver transplantation in the rat and some studies of the immunologic responses to fully allogeneic liver grafts.</a> Transplant Proc. 11, 571-577 (1979).</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=Donor+pretreatment+with+tetrahydrobiopterin+saves+pancreatic+isografts+from+ischemia+reperfusion+injury+in+a+mouse+model.">Donor pretreatment with tetrahydrobiopterin saves pancreatic isografts from ischemia reperfusion injury in a mouse model.</a> Am J Transplant. 10, 2231-2240 (2010).</li><li>Heron, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+for+accessory+cervical+heart+transplantation+in+rabbits+and+rats.">A technique for accessory cervical heart transplantation in rabbits and rats.</a> Actapathologica et microbiologica Scandinavica Section A, Pathology. 79, 366-372 (1971).</li></ol>

This study has been supported by grant #207 of the &#8220;Medizinischer Forschungsfond Tirol&#8221;, grant #2010022010 of the &#8220;MUI-START F&#246;rderungsprogramm&#8221; of the Innsbruck Medical University.

El rechazo de aloinjertos vascularizados implica una gran cantidad de diferentes medidas que sólo pueden evaluarse suficientemente utilizando modelos in vitro. Fenómenos como la sensibilización de los receptores, el procesamiento de antígenos en los órganos linfoides secundarios, sino también la diferenciación y proliferación de células inmunocompetentes son mejor caracterizados in vivo. Los modelos animales son de esta manera la herramienta ideal para la investigación traslacional 16.</...

El trasplante cardíaco representa el tratamiento de elección para los pacientes que sufren de diferentes enfermedades cardíaca terminal. Los avances en las técnicas quirúrgicas, la profilaxis más eficaz de las infecciones y nuevas pautas de inmunosupresión como resultado notablemente mejorada resultado del trasplante de órganos 6. Sin embargo, la supervivencia del injerto a largo plazo no ha mejorado rigurosamente en los últimos años 7. El rechazo crónico, que se caracteriza por la arteriosclerosis de trasplante sigue siendo un obstáculo importante para la supervivencia del injerto a largo plazo 8-11. El modelo de trasplante heterotópico de corazón en ratones proporciona una herramienta importante y válido para el análisis de mecanismo inmunológico durante aguda, así como el rechazo crónico 12-15. Hasta la fecha el modelo de trasplante más frecuente sigue siendo el abdominal trasplante de corazón de ratón utilizando la técnica de sutura. La aorta ascendente del corazón del donante se anastomosa a la unabdominal aorta y la arteria pulmonar se anastomosa a la vena cava inferior del receptor. Gran parte de la dificultad microquirúrgica del modelo de sutura se basa en el pequeño tamaño de los vasos, que se suturan 16,17. En contraste con el modelo de sutura el corazón se coloca en la región del cuello del recipiente donde la vena yugular externa se anastomosa a la arteria pulmonar y la arteria carótida común la aorta del donante. La razón de ser del desarrollo del modelo de trasplante de corazón cervical utilizando la técnica del manguito era tener un modelo animal, que permite lograr altas tasas de éxito con las habilidades básicas de microcirugía que permitirán una amplia aplicación de este modelo. Las principales ventajas de este método son la anastomosis se producen significativamente menos complicaciones tales como hemorragias y trombosis relacionada en comparación con el modelo de sutura 18.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Yasargil Clip Mini Permanent 7mm</td>
			<td>Aesculap</td>
			<td>FE720K</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro vessel clip</td>
			<td>S &#38; T</td>
			<td>B1 00396 V</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscissor</td>
			<td>FST</td>
			<td>14075-11</td>
			<td></td>
		</tr>
		<tr>
			<td rowspan="2">Vesseldilatator</td>
			<td rowspan="2">S &#38; T</td>
			<td rowspan="2">D-5a.2 00125</td>
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		<tr>
			<td></td>
		</tr>
		<tr>
			<td>Microforceps</td>
			<td>FST</td>
			<td>Dumont 11271- 30</td>
			<td></td>
		</tr>
		<tr>
			<td>Clipapplicator</td>
			<td>S &#38; T</td>
			<td>CAF-4 00072</td>
			<td></td>
		</tr>
		<tr>
			<td>Microvessel clip</td>
			<td>S &#38; T</td>
			<td>B1 00396 V</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine cervical heart transplantation model using a modified cuff technique

<html> Los modelos de ratón son de especial interés en la investigación desde una amplia variedad de anticuerpos monoclonales y comercialmente definen endogámica y las cepas knockout están disponibles para llevar a cabo mecanicista en estudios in vivo. Mientras que los modelos de trasplante de corazón utilizando una técnica de sutura se desarrolló por primera vez con éxito en ratas, la traducción a un equivalente murino utilizado igualmente generalizada nunca se logró debido a la complejidad técnica del procedimiento de microcirugía. En contraste, las técnicas del manguito no sutura, también desarrolladas inicialmente en ratas, se adaptaron con éxito para su uso en ratones de 1-3. Esta técnica de revascularización implica dos pasos principales I) eversión del buque receptor sobre un manguito de polietileno; II) tirando del buque cedente sobre el buque receptor antes evertido y manteniéndolo en su lugar con un empate circunferencial. Esto asegura una continuidad de la capa endotelial, tiempo de funcionamiento corto y muy altas tasas de permeabilidad 4. t &quot;&gt; Utilización de esta técnica para la anastomosis vascular que lleva a cabo más de 1.000 trasplantes de corazón cervicales con una tasa de éxito del 95%. Para arterial flujo de la arteria carótida común y el arco aórtico proximal se anastomosa dando como resultado una perfusión retrógrada del corazón trasplantado. Para el drenaje venoso de la arteria pulmonar del injerto se anastomosa con la vena yugular externa del receptor 5. En esto, proporcionamos detalles adicionales de esta técnica para complementar el vídeo.

La Figura 2 muestra los datos de supervivencia de los aloinjertos cardiacos representativos de donantes BALB / c después del trasplante en MHC totalmente coincidentes C57BL / 6 destinatarios. Los corazones son rechazados en el día 7 ± 1 después del trasplante. Palpación del dedo del aloinjerto trasplantado es un método sensible para detectar el curso dependiente del tiempo del rechazo de aloinjertos. Todos los trasplantes de corazón singeneicos realizadas en nuestra serie sobrevivieron indefinida...

Watch this Scientific Journal Video about Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique at JoVE.com

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

El modelo de trasplante de corazón murino de cuello uterino es muy adecuado para inmunológica así como estudios de la lesión de reperfusión de isquemia. Hemos modificado el procedimiento utilizando una técnica de manguito no sutura y realizado más de 1.000 trasplantes de éxito con este enfoque. En esto, proporcionamos detalles adicionales de esta técnica para complementar el vídeo.

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 ...

murine-cervical-heart-transplantation-model-using-modified-cuff

Research

JoVE Journal

Medicine

13.8K vistas.  Innsbruck Medical University.  El modelo de trasplante de corazón murino de cuello uterino es muy adecuado para inmunológica así como estudios de la lesión de reperfusión de isquemia. Hemos modificado el procedimiento utilizando una técnica de manguito no sutura y realizado más de 1.000 trasplantes de éxito con este enfoque. En esto, proporcionamos detalles adicionales de esta técnica para complementar el vídeo. 

Video: Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Modelo murino de Curación de Heridas

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Investigación

Medicina

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

Un modelo murino de la médula espinal cervical Lesión en estudio post-lesional Respiratorio Neuroplasticidad

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

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 ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

El trasplante de la válvula pulmonar Utilizando un modelo de ratón de heterotópico trasplante cardiaco

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

Murino Trasplante corneal: un modelo para estudiar la forma más común de trasplante de órganos sólidos

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

Heterotopic Cervical Heart Transplantation in Mice

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

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

Heterotópico cervical trasplante cardiaco en ratones

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

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Utilizando la técnica de la manga en un modelo murino de trasplante aórtico - un Video instructivo

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

Modelo murino de trasplante de páncreas mediante una técnica modificada del pun ¢ o

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

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 ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

Modelo de trasplante de aórtica cervical murino utilizando una técnica de manguito no sutura modificada

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...