The authors declare that they have no competing financial interests.

Tiere werden in einem barrierefreie Einrichtung Erreger untergebracht. Alle Tiere erhalten menschlicher Obhut in Übereinstimmung mit den &quot;Principals of Laboratory Animal Care&quot; von der National Society for Medical Research &quot;und dem&quot; Leitfaden für die Pflege und Verwendung von Labortieren &quot;der Nationalen Akademie der Wissenschaften vorbereitet formuliert und veröffentlicht von der National Institutes of Health (NIH Publikation nicht. 86-23, überarbeitet 1985). Alle Versuche werden vom Bundesministerium für Bildung, Wissenschaft und Kultur genehmigt. 1. Empfänger Vorbereitung <ol><li> Betäuben die Empfängertier mit einer im Injektion von Xylazin (5 mg / kg Körpergewicht) und Ketamin (100 mg / kg Körpergewicht). </li><li> Entfernen Sie alle Haare der seitlichen Halsbereich des Tieres und schrubben das Operationsfeld dreimal mit chlorhexdine. </li><li> Platzieren Sie die Maus in Rückenlage auf dem Operationsfeld. </li><li> Weiter machen einen Hautschnitt von der Halsschlag incision auf der rechten Unterkiefer. </li><li> Anschließend unverblümt zu mobilisieren, die rechte äußere Halsschlagader (EJV) und teilen die Zweige zwischen Ligaturen. </li><li> Dann teilen Sie die EJV zwischen Ligaturen und achten Sie auf eine ausreichende Länge zu verlassen, um den proximalen Stumpf über die Manschette Körper umzustülpen. </li><li> Nächstes entfernen Sie den rechten Lappen der Unterkieferspeicheldrüse. </li><li> Für die venöse Anastomose passieren das proximale Ende des EJV durch die Polyethylen Manschette und befestigen Sie es am Griff der Manschette mit einem Mikroklemme. </li><li> Entfernen Sie die Ligatur am Ende des Schiffes, umstülpen, das Lumen über der Manschette und fixieren Sie es mit einem 8-0 Seidenschleife (Abbildung 1a). </li><li> Anschließend durchschneiden das Recht M. sternocleidomastoideus mit bipolaren Kauter, um Zugang zu der A. carotis communis zu gewinnen. </li><li> Weiter zu mobilisieren, die Arteria carotis communis und schneiden das Schiff zwischen den Ligaturen. </li><li> Weiter zu mobilisieren, die Arteria carotis communis und schneiden das Schiff zwischen den Ligaturen. </li><li> Führen Sie die distal Ende des Schiffes durch die Manschette und fixieren Sie es mit der Arterienklemme. </li><li> Entfernen Sie die Ligatur am Ende des Schiffes und dehnen das Lumen mit Gefäß Dilatatoren. Weiter, in Analogie zu der EJV Evert die Arterie über der Manschette und fixieren Sie es mit einem 8-0 Seidenschleife. </li></ol> 2. Herz Beschaffung <ol><li> Betäuben die Spendermaus mit einer im Injektion von Xylazin (5 mg / kg Körpergewicht) und Ketamin (100 mg / kg Körpergewicht). Dann entfernen Sie alle Haare der Bauchbereich des Tieres. Schrubben das Operationsfeld drei Mal mit chlorhexdine. Nächsten Ort die Maus in Rückenlage auf dem Operationsfeld wie zuvor beschrieben. </li><li> Nach einer Mittellinie Bauchschnitt, schieben Sie der Eingeweide mit Q-Tips auf der linken Seite, um die untere Hohlvene (IVC) aus. </li><li> Injizieren 400 ul einer 1: 4 Heparin-Natrium-Lösung in den IVC für Heparinisierung. </li><li> Führen Sie eine Thorakotomie und klappen Sie den vorderen Brustwand über kranial nach gewinneness zum Herzen. </li><li> Dann entfernen Sie den Thymus und Entlüftung machen Einschnitte in der linken und der rechten oberen Hohlvene. </li><li> Anschließend versorgen die Herzretrograd mit 4 ° Celsius HTK-Lösung durch Kanülierung der Aortenbogen mit einem 27 G Spritze auf der Ebene der brachiozephalen Zweig. </li><li> Ligieren den unteren und oberen Hohlvene mit 8-0 Seide und teilen sie distal zu den Ligaturen. </li><li> Weiter teilen den Aortenbogen auf dem Niveau des vorherigen Kanülierung und sezieren die kostenlose Lungen Stamm und teilen sie so weit distal wie möglich. </li><li> Im Anschluss daran binden die Lungenvenen mit einer Schüttligation und teilen sie distal zu den Ligaturen. </li><li> Schließlich entfernen Sie das Herz aus der Entnahmestelle und speichern sie in 4 ° Celsius HTK-Lösung (Abbildung 1b). </li></ol> 3. Implantation <ol><li> Legen Sie die Herzen Transplantat im Empfängerhalsbereich in einer umgekehrten Position (Abbildung 1c </strong&gt;). </li><li> Weiter ziehen Sie den Stamm der Lungen Herz über die Vene Manschette Konstrukt des Empfängers und fixieren Sie sie mit einem Seidenschleife (Abbildung 1d). </li><li> Führen Sie die Anastomose zwischen der Aorta des Transplantats und dem umgestülpten Arterie des Empfängertieres in der gleichen Weise (Abbildung 1e). </li><li> Anschließend entfernen Sie die venöse Klemme zuerst, gefolgt von der Arterienklemme. Das Herz wird reperfundiert und beginnt innerhalb von 1-2 min zu schlagen. Während der Reperfusion zu befeuchten das Herz mit warm (35 ° Celsius) Kochsalzlösung. </li><li> Schließlich schließen Sie die OP-Wunde mit 6-0 kontinuierliche Nähte (Abbildung 1e). </li></ol> 4. Postoperative Behandlung, Endpoint <ol><li> Gib bis zu 0,3 ml physiologischer Kochsalzlösung ip postoperativ für Flüssigkeitsersatz. </li><li> Legen Sie das Tier unter einer Wärmelampe bis Erwachen aus der Narkose. </li><li> Einmal wach, bringen Sie das Tier mit dem Gehäuse Anlage, wo sie Nahrung und Wasser ad libitum erhalten. </li><li> Auf minimize Schmerzen während der ersten 7 Tage nach der Operation zu verabreichen Carprofen (4 mg / kg alle 12 Stunden subkutan (sc). Darüber hinaus geben Buprenorphin (0,1 mg / kg) unmittelbar nach der Operation alle 12 Stunden für 5 Tage. </li><li> Erhalten das Gewicht (g) jedes Empfängertier wöchentlich, um für die richtige Nahrungsaufnahme zu beurteilen. Wenn es Hinweise auf mehr als 10-15% Gewichtsverlust im Vergleich zu Gewicht bei der Operation-date, Apathie, Lähmung oder eine sehr zurückgebogen, zu opfern das Tier mit Terminal-Isofluran Inhalation vor dem Erreichen der klinischen Endpunkt. Auch das Tier zu opfern, nach dem Herzen abgelehnt wurde. </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.

Ablehnung der gefäß Allografts bringt eine Fülle von verschiedenen Schritten, die nur unzureichend mit in-vitro-Modellen bewertet werden können. Phänomene wie die Sensibilisierung von Empfängern, die Antigenprozessierung in sekundäre lymphatische Organe, sondern auch die Differenzierung und Proliferation von immunkompetenten Zellen sind dadurch besser in vivo. Tiermodelle sind damit das ideale Werkzeug für translationale Forschung 16. Mausmodelle stellen immer noch den Goldstandard i...

Herztransplantation stellt die Therapie der Wahl bei Patienten mit verschiedenen Endstadium Herzerkrankungen. Fortschritte in der chirurgischen Techniken, effektiver Prophylaxe von Infektionen und neuartige Immunsuppressiva führte zu deutlich verbesserten Ergebnis der Organtransplantation 6. Doch langfristige Transplantatüberleben wurde nicht konsequent in den letzten Jahren 7 verbessert. Chronische Abstoßung, gekennzeichnet durch Transplantat-Arteriosklerose ist weiterhin ein großes Hindernis für die langfristige Überleben des Transplantats 8-11 sein. Das Modell der heterotopen Herztransplantation bei Mäusen eine wichtige und gültige Werkzeug zur Analyse von immunologischen Mechanismus während der akuten als auch chronischen Abstoßung 12-15. Bis heute die häufigste Transplantationsmodell ist immer noch die Bauch Maus Herztransplantation mit der Nahttechnik. Die aufsteigenden Aorta des Spenderherzens an die eine Anastomosebdominal Aorta und der Lungenarterie ist an der unteren Hohlvene des Empfängers anastomosiert. Viel von der mikro Schwierigkeit des Naht Modell basiert auf der geringen Größe der Gefäße, die genäht werden 16,17 basiert. Im Gegensatz zu dem Nahtmaterial-Modell wird das Herz in den Halsbereich des Empfängers, wo die äußere Jugularvene wird zur Lungenarterie und der Arteria carotis communis die Aorta anastomosiert des Spenders angeordnet. Das Grundprinzip der Entwicklung der Halsherztransplantation Modell mit Hilfe der Manschette Technik war es, ein Tiermodell, der hohen Erfolgsraten mit grundlegenden mikrochirurgischen Fähigkeiten, die breite Anwendung dieses Modells erlauben wird ermöglicht. Die Hauptvorteile dieses Verfahrens sind die deutlich weniger auftritt Anastomose verbundenen Komplikationen wie Blutungen und Thrombosen im Vergleich zu dem Fadenmodell 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> Mausmodelle sind von besonderem Interesse in der Forschung seit einer Vielzahl von monoklonalen Antikörpern und kommerziell Inzucht definiert und Knockout-Stämme sind in-vivo-Studien Mechanismus durchzuführen. Während Herztransplantation Modelle mit einer Nahttechnik wurden zunächst erfolgreich an Ratten entwickelt wurde, war die Übersetzung in eine ebenso weit verbreitete gebrauchte murine Äquivalent nie aufgrund der technischen Komplexität der mikrochirurgischen Verfahren erreicht. Im Gegensatz dazu wurden nicht-Nahtmanschette Techniken auch bei Ratten ursprünglich entwickelt, erfolgreich für die Verwendung in Mäusen 1-3 angepasst. Diese Technik für Revaskularisierung umfasst zwei Hauptschritte I) Umstülpen des Empfängergefäßes über einen Polyethylen Manschette; II) Ziehen des abgebenden Schiffes über die früher umgestülpt Empfängergefäßes und hielt ihn an Ort und Stelle mit einer umlaufenden Krawatte. Dies gewährleistet eine Kontinuität der Endothelschicht, kurze Betriebszeit und sehr hohe Durchgängigkeitsraten 4. t &quot;&gt; Unter Verwendung dieser Technik für die vaskuläre Anastomose führten wir mehr als 1.000 zervikalen Herztransplantationen mit einer Erfolgsquote von 95%. arterielle Zufluss der Arteria carotis communis und das proximal Aortenbogen wurden anastomosiert was eine retrograde Perfusion des transplantierten Herzens. für venöse Drainage wurde die Lungenarterie des Transplantats mit der externen Halsschlagader des Empfängers 5 anastomosiert. Hier stellen wir weitere Einzelheiten dieser Technik, um das Video zu ergänzen.

Abbildung 2 zeigt repräsentative Überlebensdaten von Herzallotransplantaten von BALB / c-Spendern nach Transplantation in vollständig übereinstimm MHC C57BL / 6 Empfängern. Herzen werden an Tag 7 ± 1 nach der Transplantation abgelehnt. Finger Palpation des transplantierten Allograft ist eine empfindliche Methode, um den zeitlichen Verlauf der Transplantatabstoßung zu erkennen. Alle syngenen Herztransplantationen in unserer Serie durchgeführt lebt auf unbestimmte Zeit (&gt; 150 Tage). <p clas...

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

Die murine Gebärmutterhalskrebs Herztransplantation Modell ist für immunologische sowie Ischämie Reperfusionsschaden Studien gut geeignet. Wir modifizierten das Verfahren mit einem nicht-Nahttechnik und Manschette durchgeführt mehr als 1.000 erfolgreiche Transplantationen mit diesem Ansatz. Hier stellen wir weitere Einzelheiten dieser Technik, um das Video zu ergänzen.

Murine Cervical Herztransplantation Modell mithilfe eines modifizierten Cuff-Technik

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 Aufrufe.  Innsbruck Medical University.  Die murine Gebärmutterhalskrebs Herztransplantation Modell ist für immunologische sowie Ischämie Reperfusionsschaden Studien gut geeignet. Wir modifizierten das Verfahren mit einem nicht-Nahttechnik und Manschette durchgeführt mehr als 1.000 erfolgreiche Transplantationen mit diesem Ansatz. Hier stellen wir weitere Einzelheiten dieser Technik, um das Video zu ergänzen. 

Serie: 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.

Mausmodell der Wundheilung

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

Forschung

Medizin

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.

Murine Heterotope Heart Transplant-Technik

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 von Pulmonalklappe Mit einem Maus-Modell der heterotopen Herztransplantation

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.

Murine Hornhauttransplantation: Ein Modell, das die häufigste Form der Organtransplantation Studieren

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.

Heterotope Cervical Herztransplantation bei Mäusen

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.

Mit Hilfe der Sleeve-Technik in einem Mausmodell der Aorta Transplantation - ein Lehr-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 ...

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.

Maus-Modell für die Bauchspeicheldrüse Transplantation mit einer modifizierten Manschette-Technik

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.

Ein präklinisches Porcine-Modell der orthotopischen Herztransplantation

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.

Murine Cervical Aortic Transplantation Modell mit einer modifizierten Non-Suture Manschettentechnik

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.