The authors declare that they have no competing financial interests.

Les animaux sont logés dans une usine sans barrière de l&#39;agent pathogène. Tous les animaux reçoivent des soins humaine en conformité avec les «directeurs de laboratoire de protection des animaux&quot; formulées par la Société nationale pour la recherche médicale »et le« Guide pour le soin et l&#39;utilisation des animaux de laboratoire »préparé par l&#39;Académie nationale des sciences et publiées par le National Institutes of Health (NIH Publication no. 86-23, révisé 1985). Toutes les expériences sont approuvés par le ministère autrichien de l&#39;Education, de la Science et de la Culture. 1. bénéficiaire Préparation <ol><li> Anesthésier l&#39;animal receveur avec une injection intramusculaire de xylazine (5 mg / kg de poids corporel) et de kétamine (100 mg / kg de poids corporel). </li><li> Retirer tous les poils de la région cervicale latérale de l&#39;animal et frotter le champ opératoire trois fois en utilisant chlorhexdine. </li><li> Passer la souris dans une position couchée sur le champ opératoire. </li><li> Ensuite faire une incision de la peau de la jugulaire incision de la mandibule inférieure droite. </li><li> Par la suite, mobiliser carrément la veine jugulaire externe droite (CPA) et de diviser les branches entre ligatures. </li><li> Ensuite, divisez la CPA entre ligatures en veillant à laisser suffisamment de longueur pour retourner la souche proximale sur le corps brassard. </li><li> Ensuite, enlevez le lobe droit de la glande sous-maxillaire. </li><li> Pour l&#39;anastomose veineuse passer l&#39;extrémité proximale de la CPA par l&#39;intermédiaire du manchon de polyéthylène et le fixer à la poignée de la manchette avec un micro pince. </li><li> Retirez la ligature à l&#39;extrémité de la cuve, retourner la lumière sur la manchette et le fixer avec un 8-0 boucle de soie (figure 1a). </li><li> Par la suite transect le muscle sterno droite avec cautérisation bipolaire pour accéder à l&#39;artère carotide commune. </li><li> Suivant mobiliser l&#39;artère carotide commune et couper le navire entre les ligatures. </li><li> Suivant mobiliser l&#39;artère carotide commune et couper le navire entre les ligatures. </li><li> Passez les distal fin de la cuve par la manchette et le fixer avec la pince de l&#39;artère. </li><li> Retirer la ligature à l&#39;extrémité du récipient et dilater la lumière en utilisant dilatateurs vasculaires. Ensuite, par analogie avec la CPA, retourner la artère sur le brassard et le fixer avec une boucle 8-0 de soie. </li></ol> 2.-Cœur achats <ol><li> Anesthésier la souris donneuse avec une injection intramusculaire de xylazine (5 mg / kg de poids corporel) et de kétamine (100 mg / kg de poids corporel). Ensuite, retirer tous les poils de la région abdominale de l&#39;animal. Frotter le champ opératoire trois fois à l&#39;aide chlorhexdine. Ensuite, placez la souris en position couchée sur le champ opératoire tel que décrit précédemment. </li><li> Après une incision abdominale médiane, retirer les viscères avec Q-tips à gauche afin d&#39;exposer la veine cave inférieure (VCI). </li><li> Injecter 400 pi d&#39;une solution à 1: 4 héparine sodium dans la VCI pour héparinisation. </li><li> Effectuez une thoracotomie et plier la paroi thoracique antérieure sur crânienne à gagner seloness au cœur. </li><li> Ensuite, retirez le thymus et faire la ventilation des incisions dans la veine cave supérieure gauche et à droite. </li><li> Par la suite, perfuser le coeur d&#39;une manière rétrograde de 4 ° Celsius solution HTK par cathétérisme de la crosse aortique avec une seringue G 27 au niveau de la branche brachiocéphalique. </li><li> Ligaturer la veine cave inférieure et supérieure avec 8-0 soie et de les diviser de manière distale pour les ligatures. </li><li> Suivant diviser la crosse de l&#39;aorte au niveau de la ponction précédente et disséquer libre du tronc pulmonaire et diviser de manière distale dans la mesure du possible. </li><li> Par la suite, lier les veines pulmonaires avec une ligature en vrac et de les diviser de manière distale pour les ligatures. </li><li> Enfin, retirez le coeur de la zone donneuse et la stocker dans 4 ° Celsius solution HTK (figure 1b). </li></ol> 3 Implantation <ol><li> Placer la greffe de cœur dans la région du cou du destinataire dans une position à l&#39;envers (figure 1c </strong&gt;). </li><li> Suivant tirer le tronc pulmonaire du cœur sur la construction veine de manchette du destinataire et le fixer avec une boucle de soie (figure 1d). </li><li> Effectuez l&#39;anastomose entre l&#39;aorte de la greffe et l&#39;artère retournée de l&#39;animal receveur de la même façon (figure 1e). </li><li> Par la suite, retirer la pince veineuse en premier, suivi par la pince artérielle. Le cœur est reperfusé et se met à battre dans 1-2 min. Au cours de la reperfusion humidifier le cœur chaud (35 ° C) une solution saline. </li><li> Enfin fermer la plaie chirurgicale avec 6-0 sutures continues (Figure 1e). </li></ol> 4. Soins postopératoires, Endpoint <ol><li> Donner jusqu&#39;à 0,3 ml ip de solution saline normale après l&#39;opération pour le remplacement des liquides. </li><li> Placez l&#39;animal sous une lampe chauffante jusqu&#39;à ce que le réveil de l&#39;anesthésie. </li><li> Une fois réveillé, le réexpédier vers le centre d&#39;hébergement où ils reçoivent de la nourriture et de l&#39;eau ad libitum. </li><li> Midouleur tienne pas compte pendant les 7 premiers jours postopératoires administrer carprofène (4 mg / kg toutes les sous-cutanée de 12 h (sc). De plus, donner Buprénorphine (0,1 mg / kg) juste après l&#39;opération toutes les 12 heures pendant 5 jours. </li><li> Obtenir le poids (g) de chaque hebdomadaire animal receveur à évaluer pour l&#39;apport nutritionnel adéquat. S&#39;il existe des preuves de plus de 10-15% de perte de poids par rapport à la masse à la chirurgie jour, l&#39;apathie, paralysant ou un dos très courbé, sacrifier l&#39;animal à l&#39;aide inhalation isoflurane borne avant d&#39;atteindre le point final clinique. Aussi sacrifier l&#39;animal après que le cœur a été rejetée. </li></ol>

<ol>
	<li>Brandacher, G., 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=Tetrahydrobiopterin+compounds+prolong+allograft+survival+independently+of+their+effect+on+nitric+oxide+synthase+activity.">Tetrahydrobiopterin compounds prolong allograft survival independently of their effect on nitric oxide synthase activity.</a> Transplantation. 81, 583-589 (2006).</li><li>Schneeberger, S., 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+effect+of+secretory+leukocyte+protease+inhibitor+(SLPI)+on+ischemia/reperfusion+injury+in+cardiac+transplantation.">The effect of secretory leukocyte protease inhibitor (SLPI) on ischemia/reperfusion injury in cardiac transplantation.</a> Am J Transplant. 8, 773-782 (2008).</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=IDO+and+regulatory+T+cell+support+are+critical+for+cytotoxic+T+lymphocyte-associated+Ag-4+Ig-mediated+long-term+solid+organ+allograft+survival.">IDO and regulatory T cell support are critical for cytotoxic T lymphocyte-associated Ag-4 Ig-mediated long-term solid organ allograft survival.</a> JImmunol. 188, 37-46 (2012).</li><li>Yamashita, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+is+essential+for+and+promotes+tolerance+to+transplanted+organs.">Heme oxygenase-1 is essential for and promotes tolerance to transplanted organs.</a> FASEB J. 20, 776-778 (2006).</li><li>Kienzl, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteomic+profiling+of+acute+cardiac+allograft+rejection.">Proteomic profiling of acute cardiac allograft rejection.</a> Transplantation. 88, 553-560 (2009).</li><li>Aurora, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Registry+of+the+International+Society+for+Heart+and+Lung+Transplantation:+tenth+official+pediatric+lung+and+heart/lung+transplantation+report--2007.">Registry of the International Society for Heart and Lung Transplantation: tenth official pediatric lung and heart/lung transplantation report--2007.</a> J Heart Lung Transplant. 26, 1223-1228 (2007).</li><li>H&#228;yry, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+allograft+vasculopathy+new+strategies+for+drug+development.">Chronic allograft vasculopathy new strategies for drug development.</a> Transplant Proc. 30, 3989-3990 (1998).</li><li>Hosenpud, J. 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.

Rejet des allogreffes vascularisées entraîne une pléthore de différentes étapes qui ne peuvent être insuffisamment évalués en utilisant des modèles in vitro. Les phénomènes tels que la sensibilisation des destinataires, le traitement de l&#39;antigène dans les organes lymphoïdes secondaires, mais aussi la différenciation et la prolifération des cellules immunocompétentes sont mieux caractérisée in vivo. Les modèles animaux sont ainsi l&#39;outil idéal pour la recherche translationne...

La transplantation cardiaque représente le traitement de choix pour les patients atteints de différentes maladies cardiaques en phase terminale. Les progrès des techniques chirurgicales, la prophylaxie des infections plus efficace, et de nouveaux traitements immunosuppresseurs ont donné lieu à une amélioration des résultats nettement de la transplantation d&#39;organes 6. Cependant, la survie du greffon à long terme ne s&#39;est pas améliorée avec rigueur au cours des dernières années 7. Le rejet chronique, caractérisé par l&#39;artériosclérose de greffe continue d&#39;être un obstacle majeur à long terme la survie du greffon 8-11. Le modèle de transplantation cardiaque hétérotopique chez la souris fournit un outil important et valable pour l&#39;analyse de mécanisme immunologique pendant aiguë ainsi que le rejet chronique 12-15. A ce jour le modèle de la transplantation est le plus souvent encore la transplantation cardiaque abdominale de la souris en utilisant la technique de suture. L&#39;aorte ascendante du cœur du donneur est anastomosé à l&#39;unaorte bdominal et l&#39;artère pulmonaire est anastomosée à la veine cave inférieure du receveur. Une grande partie de la difficulté de microchirurgie du modèle de fil de suture est basée sur la taille réduite des vaisseaux, qui sont suturées 16,17. Contrairement au modèle de fil de suture est placé au coeur de la région de col du récipient où la veine jugulaire externe est anastomosé à l&#39;artère pulmonaire et l&#39;artère carotide commune de l&#39;aorte du donneur. La raison de l&#39;évolution du modèle de transplantation cardiaque col de l&#39;utérus en utilisant la technique du brassard était d&#39;avoir un modèle animal, ce qui permet d&#39;atteindre des taux de réussite élevés avec des compétences de base de microchirurgie qui permettront une large application de ce modèle. Les principaux avantages de cette méthode sont l&#39;anastomose se produit significativement moins de complications telles que hémorragie et de thrombose liée par rapport au modèle de fil de suture 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>
			<td></td>
		</tr>
		<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> Des modèles de souris sont d&#39;un intérêt particulier dans la recherche depuis une grande variété d&#39;anticorps monoclonaux et définis dans le commerce consanguine et souches de finale sont disponibles pour effectuer mécaniste des études in vivo. Alors que les modèles de transplantation cardiaque en utilisant une technique de suture ont été développés avec succès chez le rat, la traduction dans la tout aussi répandue équivalent murin utilisé n&#39;a jamais été atteint en raison de la complexité technique de la procédure microchirurgicale. En revanche, les techniques non-suture boutons de manchette, également développé initialement chez le rat, ont été adaptés avec succès pour une utilisation chez les souris 1-3. Cette technique de revascularisation comporte deux grandes étapes I) éversantes le navire bénéficiaire sur une manchette de polyéthylène; II) en tirant le navire donneur sur le navire bénéficiaire anciennement retournée et le maintenir en place avec une cravate périphérique. Ceci assure une continuité de la couche endothéliale, de courte durée de fonctionnement et les taux de perméabilité très élevée 4. t &quot;&gt; Grâce à cette technique pour l&#39;anastomose vasculaire nous avons joué plus de 1000 transplantations cardiaques col de l&#39;utérus avec un taux de réussite global de 95%. Pour l&#39;apport artériel de l&#39;artère carotide commune et la crosse de l&#39;aorte proximale ont été anastomosée résultant en une perfusion rétrograde de la greffe du coeur. Pour le drainage veineux de l&#39;artère pulmonaire de la greffe a été anastomosée avec la veine jugulaire externe du destinataire 5. Ici, nous fournissons des détails supplémentaires de cette technique pour compléter la vidéo.

La figure 2 montre les données de survie représentatives des allogreffes cardiaques provenant de donneurs BALB / c après la transplantation en pleine CMH incompatibles C57BL / 6 destinataires. Les coeurs sont rejetées au jour 7 ± 1 après la transplantation. Palpation de l&#39;allogreffe transplanté est une méthode sensible pour détecter le cours dépend de rejet d&#39;allogreffe de temps. Tous les transplantations cardiaques congénitaux réalisées dans notre série ont survécu indéfiniment ...

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

Le modèle de transplantation cardiaque col de l&#39;utérus de souris est bien adapté pour immunologique ainsi que des études de lésions d&#39;ischémie reperfusion. Nous avons modifié la procédure en utilisant une technique de manchette non-suture et effectué plus de 1.000 transplantations réussies avec cette approche. Ici, nous fournissons des détails supplémentaires de cette technique pour compléter la vidéo.

Murin col de l&#39;utérus Cœur, la transplantation en utilisant une technique de modification de manchette

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

13.7K vues.  Innsbruck Medical University.  Le modèle de transplantation cardiaque col de l'utérus de souris est bien adapté pour immunologique ainsi que des études de lésions d'ischémie reperfusion. Nous avons modifié la procédure en utilisant une technique de manchette non-suture et effectué plus de 1.000 transplantations réussies avec cette approche. Ici, nous fournissons des détails supplémentaires de cette technique pour compléter la vidéo. 

Vidéo: 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.

Modèle murin de la cicatrisation

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

Recherche

Médecine

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.

Murin de transplantation cardiaque hétérotopique 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 ...

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.

Transplantation de valve pulmonaire en utilisant un modèle de souris de transplantation cardiaque hétérotopique

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.

Murin de transplantation de la cornée: Un modèle pour étudier la forme la plus courante de la transplantation d&#39;organes solides

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.

Heterotopic col de l&#39;utérus chez la souris Transplantation cardiaque

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.

Utilisant la Technique de la manche dans un modèle murin de la greffe aortique - une vidéo d’instruction

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.

Modèle de souris pour une Transplantation du pancréas en utilisant une Technique mis à jour le brassard

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 modèle porcin préclinique de transplantation orthotopic de coeur

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.

Modèle de transplantation aortique cervicale de Murine utilisant une technique modifiée de manchette de non-suture

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

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.