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1. Donatori di funzionamento <ol><li> Anestetizzare ratti maschi peso 200 ± 20 grammi con isoflurano sul cono maschera (il 3% per l&#39;induzione, 2% durante l&#39;operazione, 1 &#39;aria l / min di flusso, FiO2 70%). </li><li> Eseguire una laparotomia mediana larga e trasversale e cauterizzare i vasi epigastrici su entrambi i lati. </li><li> Tagliare il legamento falciforme, e separare la vena diaframmatica sinistra dal suprehepatic vena cava. Infine, dividere la vena diaframmatica tra due 7-0 legature di seta. </li><li> Tagliare il triangolare di sinistra ed i legamenti gastro-epatiche. </li><li> Dividere il legamento epato-esofageo e l&#39;arteria tra 7-0 legature (coagulazione è una valida alternativa). </li><li> Isolare il infraepatica vena cava fino alla vena renale sinistra. Separare la vena renale destra dai tessuti circostanti e dividerlo tra 10-0 e 7-0 legatura, sulle sue estremità prossimale e distale, rispettivamente. </li><li> Dividere la vena surrenale destra tra 7-0 ligaturES e libera il fegato dai suoi legamenti posteriori, tagliando sotto una leggera trazione. </li><li> Isolare la vena gastrosplenico e dividerlo tra 10-0 e 7-0 legatura, sulle sue estremità prossimale e distale, rispettivamente. </li><li> Dividere l&#39;arteria epatica corretta tra 7-0 legami. </li><li> Isolare il duodeno-pancreatico vena e divisa fra un 10-0 e 7-0 uno legatura, sulla sua estremità prossimale e distale, rispettivamente. </li><li> Inserire un 22G, stent 3,5 mm nel dotto biliare comune, praticando una piccola incisione di 1 cm di distanza dal ilo epatico. Fissare la stent in posizione con una legatura 7-0. </li><li> Iniettare 10 UI di eparina, diluito in 1 ml di soluzione salina normale, attraverso la vena dorsale del pene. </li><li> Incannulare la vena porta (per quanto possibile dall&#39;ilo) con un ago 21G e delicatamente lavare il fegato con 20 ml di lattato suoneria freddo (o altre soluzioni di conservazione). Al tempo stesso, tagliare la vena cava inferiore le vene renali per consentire un adeguato outflow. Perfusione epatica freddo dovrebbe durare tra uno e due minuti. </li><li> Tagliare la vena porta al di sotto del duodeno-pancreatico vena. </li><li> Tagliare il dotto biliare comune distalmente alla legatura attorno allo stent. </li><li> Completa l&#39;incisione della vena cava infraepatica. </li><li> Tagliare il sovraepatica vena cava skim al diaframma e rimuovere il fegato. </li><li> Mettere il fegato in una bacinella piena di Ringer lattato freddo e laici del bacino su una base di ghiaccio. </li></ol> 2. Preparazione Graft <ol><li> Inserire la vena porta nel suo bracciale, poi questi vanno rovesciati con la nave intorno al bordo per esporre il intima. Bloccare il segmento di estroflesso sul bracciale con una legatura 7-0. </li><li> Ripetere la stessa operazione sul infraepatica vena cava. </li><li> Posizionare un micro-clamp (4-6 mm di lunghezza), sulla parte prossimale del sottoepatica vena cava. Questa operazione evita la perdita di sangue dopo riperfusione portale. </li><li> Capovolgere il fegato e inserire due Prolene 8-0 punti di suturai bordi laterali opposti della sovraepatica vena cava, dall&#39;esterno verso l&#39;interno. </li><li> Conservare l&#39;innesto (immerso in soluzione di Ringer lattato) in frigorifero a 0-4 ° C. </li></ol> 3. Destinatario di funzionamento <ol><li> Anestetizzare ratti maschi peso 200 ± 20 grammi con isoflurano sul cono maschera (il 3% per l&#39;induzione, 2% durante l&#39;operazione, 1 &#39;aria l / min di flusso, FiO2 70%) e mantenere su un blocco caldo. Da notare, pesi beneficiari e dei donatori dovrebbe essere abbinato (± 40 grammi). </li><li> Iniettare 10 ml di soluzione fisiologica per via sottocutanea e 0,03 g di piperacillina / tazobactam intramuscolare prima della laparotomia. </li><li> Eseguire una linea mediana xifo-pubica laparotomia. </li><li> Inverso il processo xifoideo con un auto-statici forcipe per esporre meglio il fegato. </li><li> Tagliare i legamenti anteriori e dividere la vena diaframmatica sinistra, tra due legami 7-0. </li><li> Dividere il legamento epato-esofageo e l&#39;arteria tra 7-0 legature (coagulation è una valida alternativa). Non mostrato nel video (identico al funzionamento del donatore). </li><li> Isolare il infraepatica vena cava fino alla vena renale destra. </li><li> Dividere la vena surrenale destra tra 7-0 legature e liberare il fegato dai suoi legamenti posteriori, tagliando in trazione leggera (non mostrato nel video (identico al funzionamento dei donatori)). </li><li> Posizionare un 7-0 legatura attorno al dotto biliare comune, proprio sotto la sua divisione. In altre parole legatura uno millimetri al di sotto e tagliare in mezzo. </li><li> Isolare l&#39;arteria epatica all&#39;ilo e dividerlo tra i 7-0 legature. </li><li> Separare delicatamente i rami portale di sinistra e di destra. Prestare attenzione a non danneggiare l&#39;arteria epatica comune o gastro-vena splenica. </li><li> Posizionare l&#39;anello attorno alla cava vena cava, cranialmente alla vena renale destra. Quindi fissare la parete del vaso per l&#39;anello mettendo quattro cardinali 9-0 punti di sutura. </li><li> Ripetere la stessa operazione con l&#39;anello portale sulla vena porta. </li&gt; <li> Iniettare 10 UI di eparina, diluito in 1 ml di soluzione salina normale, attraverso la vena dorsale del pene. </li><li> Impostare l&#39;anestesia 0,25% e la FiO2 al 100%. </li><li> Bloccare il infraepatica vena cava appena sopra la vena renale destra, quindi fissare il porta appena sopra la vena gastro-vena splenica. Infine bloccare la vena cava sovraepatica essere sicuri di includere almeno 2 mm di diaframma. </li><li> Tagliare il vicino sovraepatica vena cava al parenchima. </li><li> Tagliare la sinistra ei rami portale di destra e dividere il setto in mezzo. </li><li> Tagliare il vicino infraepatica vena cava al parenchima, facendo attenzione a non incidere uno dei punti di sutura precedentemente posizionati. Rimuovere il fegato e posizionare l&#39;innesto nella cavità addominale. </li><li> Eseguire un end-to-end anastomosi tra la vena cava sovraepatica del graft e del destinatario. Iniziare con la sutura continua della parete posteriore da sinistra a destra, quindi procedere sulla parete anteriore da destra verso left Mantenere la sutura anteriore sciolto fino alla fine per consentire un lavaggio generosa del lume interno prima di serrare le estremità. </li><li> Inserire il ramo caudale del approssimatore nel manico anello portale, quindi inserire la vena porta dell&#39;innesto nella fessura del ramo craniale del approssimatore, con il polsino portale trovando la sua posizione nella fessura. </li><li> Chiudere la approssimatore durante il lavaggio del destinatario vena porta con soluzione fisiologica e consentire il bracciale da inserire nel vaso. Fissare il destinatario vena porta attorno al suo polso con un pareggio 7-0 circonferenziale. </li><li> Rimuovere il approssimatore. Aprire il morsetto sul sovraepatica vena cava e vena porta al fine di: flusso portale è ora ristabilita. </li><li> Rimuovere l&#39;anello portale tagliando i 9-0 legature con olio extra-fini forbici. </li><li> Inserire il ramo caudale della approssimatore nel manico ad anello caval, quindi inserire il infraepatica vena cava del trapianto nella fessura del cranio<html> ramo del approssimatore, con il polsino cava trovando la sua posizione nella fessura. </li><li> Chiudere la approssimatore durante il lavaggio del destinatario sottoepatica vena cava con soluzione fisiologica e consentire il bracciale da inserire nel vaso. Fissare il destinatario vena cava attorno al suo polso con un pareggio 7-0 circonferenziale. </li><li> Rimuovere il approssimatore. Aprire le microclamps sul infraepatica vena cava dell&#39;innesto e creditore in ordine: il flusso venoso è ora ristabilita. </li><li> Eseguire una incisione parziale del dotto biliare comune del destinatario, applicare una leggera trazione verso la testa, e inserire il lato libero dello stent precedentemente inserita nel dotto biliare dell&#39;innesto. Fissare lo stent con un pareggio 7-0 circonferenziale. </li><li> Chiudere l&#39;animale da strati con 5-0 punti di sutura. Lasciare l&#39;acqua libera e alimenti provenienti da risveglio. </li><li> Amministrare analgesia efficace secondo i protocolli locali istituzionali. Una durata minima di 24 h è raccomandato. </li></ol>_content &quot;&gt; Da notare: end-to-end anastomosi dell&#39;arteria epatica può essere eseguita, quando richiesto dal disegno sperimentale (l&#39;arteria non è necessario per sopravvivenza a lungo termine dopo trapianto di fegato di ratto). 4. Risultati rappresentativi La tecnica descritta può essere masterizzato con successo dal nostro team, dopo circa 10 interventi di formazione. Ha permesso al 100% a lungo termine (&gt; 21 giorni) la sopravvivenza a 36 consecutive Lewis-to-Lewis e scuri Agouti-to-Lewis trapianti, dopo il periodo di formazione breve (un certo numero di destinatari singenici trapianto di fegato sono stati tenuti in vita per oltre 12 da mesi). Post-trapianto singenici giorno 1 test di funzionalità epatica ha mostrato mediana aspartato transaminasi (AST): 84 U / l (51,7-92,7) e alanina mediana transaminasi (ALT): 172,5 U / l (127,5-240,2). La fase anepatica media (dalla vena porta di bloccaggio ad innesto re-perfusione) è stata del 14 ± 2 minuti, con la più lunga fase è la vena cava sovraepatica anastomosi (in esecuzione sFUTURO) Tempo di 9 ± 2 min. Il rapido linker sistema ha permesso il posizionamento di 1,55 mm-foro manette vena porta e 2,40 mm a canna manette vena cava (destinatari peso 200 ± 20 g). Al contrario, quando abbiamo provato il trapianto usando la tecnica Kamada, essi non potevano di 1,40 mm e 2,16 millimetri, rispettivamente (ratti corrispondenti per tensione e peso). Come mostrato in Figura 7, rapido-linker anelli sono progettati per mantenere destinatario su navi ottimale stiramento, permettendo minimo spreco pinza e lunghezza, che si traduce in risultati emodinamici più a-fisiologiche. <img src="/files/ftp_upload/4143/4143fig1.jpg" alt="Figura 1" /> Figura 1. Misure Anelli. <img src="/files/ftp_upload/4143/4143fig2.jpg" alt="Figura 2" /> Figura 2. Misure Microclamps. <img src="/files/ftp_upload/4143/4143fig3.jpg" alt="Figura 3" /> Figura 3. Quick-linker kit. <img src="/files/ftp_upload/4143/4143fig4.jpg" alt="Figura 4" /> Figura 4. Quick-linker armata. <img src="/files/ftp_upload/4143/4143fig5.jpg" alt="Figura 5" /> Figura 5. Quick-linker chiuso. <img src="/files/ftp_upload/4143/4143fig6.jpg" alt="Figura 6" /> Figura 6. Morsetto SHVC. <img src="/files/ftp_upload/4143/4143fig7.jpg" alt="Figura 7" /> Figura 7. Classic vs quick-linker assistita bracciale-anastomosi emodinamica. <img src="/files/ftp_upload/4143/4143fig8.jpg" alt="Figura 8" /> Figura 8. Donatore e la preparazione della cute del destinatario e incisione.

<ol>
	<li>Starzl, T. E., Marchioro, T. L., Porter, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+and+clinical+observation+after+homotransplantation+of+the+whole+liver.">Experimental and clinical observation after homotransplantation of the whole liver.</a> Revue internationale d'hepatologie. 15, 1447 (1965).</li><li>Starzl, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+liver+transplantation.">Evolution of liver transplantation.</a> Hepatology. 2, 614S-636S (1982).</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 (1979).</li><li>Kashfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplantation proceedings. 37, 185-188 (2005).</li><li>Hori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+and+innovative+techniques+for+liver+transplantation+in+rats:+A+surgical+guide.">Comprehensive and innovative techniques for liver transplantation in rats: A surgical guide.</a> World Journal of Gastroenterology: WJG. 16, 3120 (2010).</li><li>Oldani, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+technique+for+rat+liver+transplantation+using+Quick+Linker+system:+a+preliminary+result.">A novel technique for rat liver transplantation using Quick Linker system: a preliminary result.</a> Journal of Surgical Research. 149, 303-309 (2008).</li><li>Nosaka, T., Bowers, J. L., Cay, O., Clouse, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biliary+complications+after+orthotopic+liver+transplantation+in+rats.">Biliary complications after orthotopic liver transplantation in rats.</a> Surg. Today. 29, 963-965 (1999).</li><li>Li, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+incidence+of+biliary+complications+in+rat+liver+transplantation:+can+we+avoid+it.">High incidence of biliary complications in rat liver transplantation: can we avoid it.</a> World J. Gastroenterol. 17, 3140-3144 (2011).</li><li>Peng, Y., Gong, J. P., Yan, L. N., Li, S. B., Li, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+two-cuff+technique+for+orthotopic+liver+transplantation+in+rat.">Improved two-cuff technique for orthotopic liver transplantation in rat.</a> Hepatobiliary Pancreat. Dis. Int. 3, 33-37 (2004).</li><li>Urata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+survival+in+rat+liver+transplantation+with+extended+cold+preservation:+role+of+portal+vein+clamping+time.">Decreased survival in rat liver transplantation with extended cold preservation: role of portal vein clamping time.</a> Hepatology. 28, 366-373 (1998).</li></ol>

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L&#39;attuale modello di ratto trapianto di fegato può essere stabilita facilmente. Nella nostra esperienza, alcuni punti chiave ridurre al minimo il rischio di morte (≤ 20%). Morti precoci, che si verificano entro le prime tre ore da riperfusione, sono spesso causa di un infarto intestinale, come conseguenza di un portale lungo bloccaggio. Si raccomanda pertanto di mantenere la fase anepatica entro il limite ideale di 15 minuti (20 minuti ancora accettabile). Central embolia gassosa può anche essere una causa di morte improvvisa ed è obbligatoria per scaricare tutte le vene prima di anastomosi. I decessi che si verificano tra il giorno post-operatorio uno e tre sono spesso dovute ad insufficienza epatica o trombosi della vena cava. Bisogna dunque evitare qualsiasi innesto aggressiva lavaggio prima della espianto nel donatore (che a filo con 20 ml in circa 60 secondi) e garantire una conservazione adeguata freddo (full immersion in 0-4 ° C soluzione di Ringer lattato). Il rischio di trombosipuò essere diminuito minimizzando la manipolazione dell&#39;intima e preservare l&#39;integrità. Si sconsiglia l&#39;uso di terapia anticoagulante dopo l&#39;intervento chirurgico. Morti tra il giorno post-operatorio quattro e nove sono spesso legati alla colangite batterica 7. Sulla base della nostra esperienza, una singola dose di piperacilline / tazobactam (0,03 g) data prima della laparotomia è sufficiente per ridurre al minimo il rischio di infezione dotto biliare comune. La morte è un evento raro dopo giorno 10. Tuttavia, a causa della mancanza di flusso arterioso, alcuni problemi dotto biliare può essere osservato con cholastasis 8. A seconda delle esigenze del ricercatore uno arterializzazione può essere considerato 9. Da notare, un tempo di ischemia fredda fino a 24 ore può essere considerato accettabile 10. Il rapido linker sistema è stato originariamente progettato per eseguire il trapianto di fegato con tre gemelli (anastomosi fase anepatica molto breve). Tuttavia, a tre bracciale TEchnique richiede l&#39;uso di pinze molto piccole per la sovraepatica vena cava (SHVC) bracciale e non è stato ampiamente accettato a causa del rischio di problemi al fegato deflusso 11-14. Abbiamo quindi favorire una rapida linker assistito a tre bracciale tecnica solo quando i tempi di ischemia calda inferiori a dieci minuti sono obbligatori. Nella maggior parte degli studi tuttavia, fasi di impianto fino a 15-18 minuti sono accettabili, consentendo l&#39;uso di due bracciale tecnica, e un drenaggio più fisiologico SHVC. Come mostrato in questo documento video, si usa abitualmente il rapido-linker sistema nella seconda parte del solo impianto. Mentre po &#39;di tempo chirurgico è necessario per posizionare le maniglie quick-linker, questa parte del procedimento è associato a rischi minimi e permette una riduzione dei critici fase anepatica grazie alla maggiore facilità di inserimento polsini. Altri vantaggi del sistema rapido-linker sono meglio un innesto-destinatario allineamento nave e l&#39;uso di più ampi polsini con risultati migliori rispetto al emodinamicitecniche precedentemente descritte (compreso quello descritto da Kamada et al.) 6 (Fig. 7).

<table border="1"><tbody><tr><td> Tool / Reagente </td><td> Azienda </td><td> Catalogo n. </td><td> Commento </td></tr><tr><td> Bracciale Portal </td><td> <a href="http://altecweb.com" target="_blank">altecweb.com</a> o <a href="http://smiths-medical.com" target="_blank">fabbri-medical.com</a> </td><td> Coil 01-96-1729 800/100/420 </td><td> Bore: 1,55 millimetri Muro: 0.30mm Lunghezza: 3,5 mm </td></tr><tr><td> V bracciale infraepatica cava </td><td> <a href="http://altecweb.com" target="_blank">altecweb.com</a> o <a href="http://smiths-medical.com" target="_blank">fabbri-medical.com</a> </td><td> Coil 01-96-1733 800/100/540 </td><td> Bore: 2,40 millimetri Muro: 0.30mm <br/&gt; Lunghezza: 3,5 mm </td></tr><tr><td> Stent dotto biliare comune </td><td> comune 22G venflon </td><td></td><td></td></tr><tr><td> 7-0 cravatte di seta </td><td> <a href="http://georgetiemann.com" target="_blank">georgetiemann.com</a> </td><td> 160-1226-7 / 0 </td><td></td></tr><tr><td> 10-0 prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> 9-0 prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> 8-0 prolene </td><td> <a href="http://ethicon.com" target="_blank">ethicon.com</a> </td><td></td><td></td></tr><tr><td> Dritti pinze micro </td><td> <a href="http://s-and-t.net" target="_blank">s-e-t</a>. Net </td><td> FRC-15RM-8 </td><td></td></tr><tr><td> Curve micro pinza </td><td> <a href="http://s-and-t.net" target="_blank">s-e-t.net</a> </td><td> FRS-15-RM-8 </td><td></td></tr><tr><td> Dissection porta-aghi </td><td> <a href="http://s-and-t.net" target="_blank">s-e-t.net</a> </td><td> B-15-8,2 </td><td></td></tr><tr><td> Anastomosi porta-aghi </td><td> <a href="http://rumex.net" target="_blank">rumex.net</a> </td><td> 8-021T </td><td></td></tr><tr><td> Morsetti Micro </td><td> <a href="http://finescience.com" target="_blank">finescience.com</a> </td><td> 18055-03 </td><td> 6 x 1mm </td></tr><tr><td> Micro morsetti applicatore </td><td> <a href="http://finescience.com" target="_blank">finescience.com</a> </td><td> 18057-14 </td><td></td></tr&gt; <tr><td> Sovraepatica v cava morsetto </td><td> <a href="http://pharmap.ch" target="_blank">pharmap.ch</a> </td><td> 13.349.14 </td><td></td></tr><tr><td> Quick-linker anelli </td><td> <a href="http://dltchiropody.co.uk" target="_blank">dltchiropody.co.uk</a> </td><td> PAR 10 </td><td> modificati lame di bisturi </td></tr><tr><td> Quick-Linker approssimatore </td><td> <a href="http://pharmap.ch" target="_blank">pharmap.ch</a> </td><td> UD-04-275 </td><td> pinza Kocher modificata di </td></tr></tbody></table>

orthotopic liver transplantation in rats

Progressi clinici nel campo del trapianto di fegato è stata ampiamente supportata da 1,2 modelli animali. Dopo la pubblicazione del primo trapianto ortotopico di fegato di ratto nel 1979 da Kamada et al. 3, questo modello è rimasto il gold standard, nonostante le varie tecniche proposte alternative 4. Tuttavia, il suo utilizzo più ampio è limitato dalla sua curva di apprendimento ripida 5. In questo documento video, mostrano una revisione semplice e facile da stabilire-di Kamada a due bracciale tecnica. La vena cava sovraepatica anastomosi viene eseguita manualmente con una sutura continua, e la vena porta e infraepatica anastomosi vena cava vengono eseguite utilizzando un sistema di quick-linker bracciale 6. Produzione del quick-linker kit è indicato in un documento video separato.

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

Orthotopic Liver Transplantation in Rats

Vi presentiamo uno facile da stabilire revisione della classica due-bracciale tecnica di trapianto di fegato nel ratto.

Trapianto di fegato in ratti

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

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29.7K Views.  University of Geneva Hospitals. Vi presentiamo uno facile da stabilire revisione della classica due-bracciale tecnica di trapianto di fegato nel ratto.

Video: Orthotopic Liver Transplantation in Rats

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

Sviluppo di bronchiolite obliterante in un modello murino di trapianto polmonare ortotopico

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

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Orthotopic Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

Ortotopico di intestino tenue di ratti

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

Aortica trapianto ortotopico in Mouse per lo studio della malattia vascolare

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Procedure chirurgiche per un modello di Rat parziale trapianto di fegato con la ricostruzione arteriosa epatica

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

Tecnica di Suini appalti Fegato e trapianto ortotopico con un attivo Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

Ortotopico degli arti posteriori del trapianto di mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

Un modello del ratto di orthotopic del trapianto del fegato usando una tecnica novella dell'anastomosi magnetico per la ricostruzione della Vena Cava Suprahepatic

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

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 modello pre-clinico di trapianto di cuore ortotopico

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

Complicazioni ridotte dopo la riconnessione arteriosa in un modello di ratto di trapianto di fegato ortotopico

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Left Lung Orthotopic Transplantation in a Juvenile Porcine Model for ESLP

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. However, lung transplantation is limited by a shortage of available donor organs. As such, there is high waitlist mortality. Ex situ lung perfusion (ESLP) has increased donor lung utilization rates in some centers by 15%-20%. ESLP has been applied as a method to assess and recondition marginal donor lungs and has demonstrated acceptable short- and long-term outcomes following transplantation of extended criteria donor (ECD) lungs. Large animal (in vivo) transplantation models are required to validate ongoing in vitro research findings. Anatomic and physiologic differences between humans and pigs pose significant technical and anesthetic challenges. An easily reproducible transplant model would permit the&#160;in vivo&#160;validation of current ESLP strategies and the preclinical evaluation of various interventions designed to improve donor lung function. This protocol describes a porcine model of orthotopic left lung allotransplantation. This includes anesthetic and surgical techniques, a customized surgical checklist, troubleshooting, modifications, and the benefits and limitations of the approach.

This protocol describes a juvenile porcine model of orthotopic left lung allotransplantation designed for use with ESLP research. Focus is made on anesthetic and surgical techniques, as well as critical steps and troubleshooting.

Trapianto ortotopico di polmone sinistro in un modello suino giovane per ESLP

Lung transplantation is the gold-standard treatment for end-stage lung disease, with over 4,600 lung transplantations performed worldwide annually. ...