1. Donatori di funzionamento <ol><li> Il topo donatore deve essere tenuto a digiuno per 24 ore (libero accesso all&#39;acqua / soluzione di glucosio). </li><li> Per indurre l&#39;anestesia isoflurano inalazione, inizia con il 2% di atomizzatore standard, e quindi ridurre a 1% dopo aver eseguito laparotomia. Eseguire un pizzico punta per controllare la sedazione. </li><li> Shave l&#39;addome e pulire con la preparazione della pelle 3 volte (Kodan). Quindi, eseguire una incisione mediana dopo somministrazione sottocutanea di analgesico. </li><li> Dopo l&#39;innesto è avvolto in soluzione salina garza imbevuta, separare le aderenze fisiologiche tra il pancreas e il colon ascendente delicatamente con un Q-tip (sotto il microscopio operatorio con un ingrandimento 6x). </li><li> Legare e sezionare i vasi coliche ileocecale e destra e centro con 7-0 seta. Dopo che il colon ascendente si sviluppa sul lato destro della vena mesenterica superiore (SMV), i vasi coliche ileocecale, destro e centrale può essere identificato per la legatura e la divisione con legami 7-0. </li><li> Disimpegno stomaco verso l&#39;alto, in modo che l&#39;intera SMV è raddrizzato ed esposta. Utilizzare un morsetto zanzara per lo svincolo. La pinza è detenuto da massa plastilina formata in forma, se necessario. </li><li> Legare e dividere i pancreatico-duodenali vene provenienti dalla SMV. Tutti piccole pancreatico-duodenale vene provenienti dal SMV deve essere accuratamente identificata, ligato con 7-0 seta e divisa prima che il tessuto pancreatico può essere rimosso dal trapianto. </li><li> Legare e sezionare il tessuto connettivo perde compresi tutti linfatici tra la SMV e l&#39;aorta addominale. Con l&#39;innesto ancora sul lato destro dell&#39;addome, il tessuto connettivo perde compresi tutti linfatici tra la SMV e l&#39;aorta addominale è accessibile. Questo tessuto connettivo deve essere ligati utilizzando 7-0 seta e diviso per evitare lymphorrhea postoperatoria dal graft intestinale. </li><li> Legare e sezionare l&#39;arteria renale destra. Dopo che il tessuto connettivo è diviso, l&#39;arteria renale destra diventa accessible ed è legata e sezionata utilizzando seta 7-0. </li><li> Sistemica heparinise il topo con 200 unità di eparina iv attraverso la vena del pene. </li><li> Legare le arterie marginali, e dividere il piccolo intestino al duodeno-digiunale giunzione e l&#39;ileo terminale. </li><li> L&#39;aorta è ligato prossimalmente all&#39;origine della SMA. La vena porta è attraversata alla confluenza con la vena splenica. Poi l&#39;innesto viene raccolto con il suo peduncolo vascolare che consiste della SMA con un segmento aortico. </li></ol> 2. Procedura Backtable <ol><li> Perfondere l&#39;innesto con 3 ml di soluzione University of Wisconsin (UW) a 4 ° C tramite il condotto aortica e irrigare il lume intestinale dall&#39;estremità digiunale con 30 ml di soluzione Nebacetin Uro-N a 4 ° C (irrigazione lumen è obbligatorio) . </li></ol> Immediatamente dopo l&#39;estrazione della protesi, il condotto aortica è utilizzato per perfusione con 3 ml di soluzione refrigerata UW. Per questo, un catetere 20 G iv sullauna siringa da 10 ml viene utilizzato. Il perfusato deve essere osservato defluire liberamente dalla vena porta divisa. Per l&#39;irrigazione intestinale con Nebacetin, una siringa da 50 ml viene utilizzato. <ol start="2"><li> Memorizzare l&#39;innesto in soluzione UW a 4 ° C durante la preparazione del destinatario. </li></ol> 3. Destinatario di funzionamento <ol><li> Il ratto del destinatario deve essere tenuto a digiuno per 24 ore (libero accesso all&#39;acqua / soluzione di glucosio). </li><li> Per indurre l&#39;anestesia isoflurano inalazione, inizia con il 2% di atomizzatore standard, e quindi ridurre a 1% dopo aver eseguito laparotomia. Eseguire un pizzico punta per controllare la sedazione. </li><li> Shave l&#39;addome e pulire con la preparazione della pelle 3 volte (Kodan). Quindi, eseguire una incisione mediana dopo la somministrazione sottocutanea di Carprofen 5 mg / kg sc per analgesia intraoperatoria. </li><li> Avvolgere l&#39;intestino destinatario in soluzione salina normale garza imbevuta e posizionarlo sul petto del destinatario. </li><li> Aprire il retroperitoneo senza mezzi termini con Q-tips, ed esporre la addominalinal aorta e vena cava inferiore appena sotto i vasi renali fino al livello dei vasi iliaci, se necessario anche utilizzare microforbici. Legare le tribuaries piccole e spinali lombari dal aorta e vena cava con 7-0 di seta per evitare la perdita di sangue. (Per fare questo, modificare lo zoom a 16x microscopio chirurgico.) </li><li> Cross-clamp l&#39;aorta e l&#39;IVC sotto i vasi renale sinistra prossimalmente e sopra la biforcazione iliaca distalmente con clips microcircolo. Un solo morsetto viene utilizzato prossimale e distale per serrare contemporaneamente entrambe le navi. Incidere entrambe le navi anteriormente con un microknife e lavare il sangue residuo. </li><li> Creare un end-to-side anastomosi tra il segmento aortico sottorenale innesto e del destinatario utilizzando un continuo 10-0 sutura Prolene .. Inizialmente, l&#39;innesto è posto sul lato destro dell&#39;addome (la testa del ratto posizionato a ore 12) per eseguire le maglie parete posteriore della anastomosi arteriosa e legatura della sutura soggiorno inferiore. Then, l&#39;innesto è rivolto verso il lato sinistro dell&#39;addome (la testa del ratto posizionato a ore 9) per esporre e suturare la parete anteriore della anastomosi. </li><li> Un end-to-side anastomosi tra la vena porta innesto e IVC del destinatario viene eseguita similmente eseguendo usando suture 10-0 Prolene. Con il topo ancora sdraiato lateralmente (testa in posizione ore 9), l&#39;innesto è posizionato sul lato sinistro dell&#39;addome e una sutura soggiorno inferiore è posto. L&#39;anastomosi viene avviata con la parete posteriore dall&#39;interno del recipiente. Dopo la sutura soggiorno inferiore è legato, i punti della parete anteriore può essere eseguita dall&#39;esterno. </li><li> Rimuovere i morsetti distali primi, seguiti da i morsetti superiori. Qualsiasi sanguinamento anastomotica è controllata dalla pressione diretta con Q-tips. L&#39;innesto deve essere controllata per riperfusione uguali e veloce. </li><li> Resecare tenue del destinatario intero dopo la legatura dei vasi mesenterici. Destinatari sottoposti enterectomia subtotale, conservando 2-3 cm di proximal digiuno e 1 cm di ileo distale. </li><li> Ripristinare la continuità enterica da prossimale (jejuno-digiunostomia) e distale (ileo-ileale) end-to-end anastomosi intestinali utilizzando un interrotto uno strato di 6-0 Monocryl sutura. Circa 16 punti di sutura sono necessari per completare le anastomosi. </li><li> Irrigare la cavità peritoneale con soluzione fisiologica fino a completa pulizia. Somministrare 2 ml di soluzione salina normale per via intraperitoneale per la sostituzione del fluido. Quindi chiudere l&#39;addome con una sutura continua con Vicryl 3-0 per lo strato muscolare più una sutura della pelle continua. </li><li> Nel periodo post-operatorio i topi dovrebbero essere tenuti a digiuno (con accesso ad acqua e una soluzione di glucosio) per un altro 24 h quindi riavviato sul ratto chow standard e acqua ad libitum. Analgesia con carprofen deve essere somministrato per 3 giorni (vedi dosi di seguito). </li></ol>

<ol>
	<li>Abu-Elmagd, K. 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=Five+Hundred+Intestinal+and+Multivisceral+Transplantations+at+a+Single+Center:+Major+Advances+With+New+Challenges.">Five Hundred Intestinal and Multivisceral Transplantations at a Single Center: Major Advances With New Challenges.</a> Ann. Surg. 250 (4), 567-56 (2009).</li><li>Berney, T., 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=Portal+versus+systemic+drainage+of+small+bowel+allografts:+comparative+assessment+of+survival,+function,+rejection,+and+bacterial+translocation.">Portal versus systemic drainage of small bowel allografts: comparative assessment of survival, function, rejection, and bacterial translocation.</a> J. Am. Coll. Surg. 195 (6), 804 (2002).</li><li>Fishbein, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+transplantation.">Intestinal transplantation.</a> N. Engl. J. Med. 361 (10), 998 (2009).</li><li>Hernandez, F., 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=Is+portal+venous+outflow+better+than+systemic+venous+outflow+in+small+bowel+transplantation?+Experimental+study+in+syngeneic+rats.">Is portal venous outflow better than systemic venous outflow in small bowel transplantation? Experimental study in syngeneic rats.</a> J. Pediatr. Surg. 40 (2), 336 (2005).</li><li>Monchik, G. 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=Transplantation+of+small+bowel+in+the+rat:+technical+and+immunological+considerations.">Transplantation of small bowel in the rat: technical and immunological considerations.</a> Surgery. 70 (5), 693 (1971).</li><li>Murase, N., 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=Graft-versus-host+disease+after+brown+Norway-to-Lewis+and+Lewis-to-Brown+Norway+rat+intestinal+transplantation+under+FK506.">Graft-versus-host disease after brown Norway-to-Lewis and Lewis-to-Brown Norway rat intestinal transplantation under FK506.</a> Transplantation. 55 (1), 1 (1993).</li><li>Murase, N., 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=Immunomodulation+of+intestinal+transplant+with+allograft+irradiation+and+simultaneous+donor+bone+marrow+infusion.">Immunomodulation of intestinal transplant with allograft irradiation and simultaneous donor bone marrow infusion.</a> Transplant Proc. 31 (1-2), 565 (1999).</li><li>Pech, T., 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=Perioperative+infliximab+application+ameliorates+acute+rejection+associated+inflammation+after+intestinal+transplantation.">Perioperative infliximab application ameliorates acute rejection associated inflammation after intestinal transplantation.</a> Am. J. Transplant. 10 (11), 2431 (2010).</li><li>Schaefer, N., 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=Resident+macrophages+are+involved+in+intestinal+transplantation-associated+inflammation+and+motoric+dysfunction+of+the+graft+muscularis.">Resident macrophages are involved in intestinal transplantation-associated inflammation and motoric dysfunction of the graft muscularis.</a> Am J Transplant. 7 (5), 1062 (2007).</li><li>Schaefer, N., 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=Mechanism+and+impact+of+organ+harvesting+and+ischemia-reperfusion+injury+within+the+graft+muscularis+in+rat+small+bowel+transplantation.+Transplant.">Mechanism and impact of organ harvesting and ischemia-reperfusion injury within the graft muscularis in rat small bowel transplantation. Transplant.</a> Proc. 38 (6), 1821 (2006).</li><li>Schaefer, N., 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=Acute+rejection+and+the+muscularis+propria+after+intestinal+transplantation:+the+alloresponse,+inflammation,+and+smooth+muscle+function.">Acute rejection and the muscularis propria after intestinal transplantation: the alloresponse, inflammation, and smooth muscle function.</a> Transplantation. 85 (10), 1465 (2008).</li><li>Xue, L., 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=Surgical+experience+of+refined+3-cuff+technique+for+orthotopic+small-bowel+transplantation+in+rat:+a+report+of+270+cases.">Surgical experience of refined 3-cuff technique for orthotopic small-bowel transplantation in rat: a report of 270 cases.</a> Am. J. Surg. 198 (1), 110 (2009).</li><li>Zhang, X. Q., 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=Simplified+techniques+in+rat+heterotopic+small+bowel+transplantation.">Simplified techniques in rat heterotopic small bowel transplantation.</a> Transplant. Proc. 38 (6), 1840 (2006).</li><li>Zhao, 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=The+establishment+of+a+new+en+bloc+combined+liver-small+bowel+transplantation+model+in+rats.+Transplant.">The establishment of a new en bloc combined liver-small bowel transplantation model in rats. Transplant.</a> Proc. 39 (1), 278 (2007).</li></ol>

Nessun conflitto di interessi dichiarati.

Mentre i modelli di trapianto intestinale nel ratto sono stati descritti già nel 1970ies 5 più dei modelli di recente attività subordinate coinvolgere eterotopico trapianto di intestino con tecniche diverse 13. Mentre le procedure eterotopiche in generale hanno il vantaggio di tecniche microchirurgiche più facile accessibilità dell&#39;innesto di valutazione, eterotopico trapianto di intestino ha il grande svantaggio di non prendere in considerazione le interazioni multiple del piccolo intesti...

<table border="1"><tbody><tr><td> Nome del reagente </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti (opzionale) </td></tr><tr><td> Università del Wisconsin (UW) soluzione (ViaSpan) </td><td> Bristol-Myers Squibb </td><td></td><td></td></tr><tr><td> Uro-Nebacetin N soluzione </td><td> Nycomed </td><td> 6967855 </td><td></td></tr><tr><td> Ampicillin </td><td> Ratiopharm </td><td></td><td></td></tr><tr><td> Carprofen (Rimadyl) </td><td> Pfizer </td><td></td><td></td></tr><tr><td> Prolene 10-0 sutura unresorbable </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Monocryl 6-0 sutura riassorbibile </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Vicryl 3-0 sutura riassorbibile </td><td> Ethicon </td><td></td><td></td></tr><tr><td> iv catetere G 20 millimetri 1.1x33 </td><td> Braun </td><td></td><td></td></tr><tr><td> iv catetere G 22 0.9x25 millimetro </td><td> Braun </td><td></td><td></td></tr><tr><td> Kodan pelle Prep </td><td> Schülke </td><td></td><td></td></tr><tr><td> NaCl soluzione di infusione 0,9% </td><td> Braun </td><td></td><td></td></tr><tr><td> Pinze curve piccolo </td><td> FineScienceTools </td><td> 11009-13 </td><td></td></tr><tr><td> Micro pinza curva </td><td> AESCULAP </td><td> BD 333 </td><td></td></tr><tr><td> Micro pinza curva </td><td> AESCULAP </td><td> FD281R </td><td></td></tr><tr><td> Micro pinza dritto 1 </td><td> WPI </td><td> 5 </td><td></td></tr><tr><td> Micro pinza diritto 2 </td><td> WPI </td><td> 2 </td><td></td></tr><tr><td> Micro supporto dell&#39;ago </td><td> WPI </td> &lt;td&gt; 14081 </td><td></td></tr><tr><td> Micro forbici </td><td> FineScienceTools </td><td> 15006-09 </td><td></td></tr><tr><td> Micro bisturi </td><td> MANI </td><td> Oftalmica coltello </td><td></td></tr><tr><td> Morsetti Micro </td><td> AESCULAP </td><td> FB329R </td><td></td></tr></tbody></table>

orthotopic small bowel transplantation in rats

Trapianto di intestino tenue è diventata un&#39;opzione accettata clinico per i pazienti con sindrome dell&#39;intestino corto e il fallimento della nutrizione parenterale (irreversibile insufficienza intestinale). In centri specializzati migliorato operativa e le strategie di gestione hanno portato a eccellenti a breve e medio termine paziente e la sopravvivenza del trapianto con una qualità della vita elevata 1,3. A differenza del più comune di trapianto di altri organi solidi (cuore cioè, fegato) molti meccanismi alla base della funzionalità del trapianto e alterazioni immunologiche indotte dal trapianto di intestino non sono del tutto noti 6,7. Gli episodi di rigetto acuto, sepsi e insufficienza cronica del trapianto sono i principali ostacoli che contribuiscono ancora meno favorevole esito a lungo termine e ostacolare un impiego più diffuso della procedura, nonostante un numero crescente di pazienti in nutrizione parenterale domiciliare che avrebbe potenzialmente beneficiare di un tale trapianto. Il piccolo intestino contieneun gran numero di leucociti passeggeri comunemente indicato come parte del sistema linfoide associato intestino (GALT), essendo questa parte del motivo della immunogenity alta del graft intestinale. La presenza e la vicinanza di molti commensali e patogeni nell&#39;intestino spiega la gravità degli episodi di sepsi volta innesto integrità della mucosa è compromessa (per esempio rigetto). Per far avanzare il campo di dati intestinali e trapianto multiorgano più generati da modelli animali affidabili e fattibile è necessario. Il modello fornito combina qui sia l&#39;affidabilità e la fattibilità, una volta stabiliti in modo standardizzato e in grado di fornire informazioni preziose nei sottostanti complessi molecolari, meccanismi cellulari e funzionali che sono attivati ​​da trapianto di intestino. Abbiamo utilizzato con successo e raffinato la procedura descritta su più di 5 anni nel nostro laboratorio 8-11. Il JoVE video-formato è particolarmente utile per dimostrare la procedura complessaDure e evitare le insidie ​​iniziali per i gruppi che intendono stabilire un modello ortotopico roditore indagando trapianto di intestino.

Normale decorso post-operatorio Gli animali trapiantati dovrebbero recuperare rapidamente dalla procedura di cui una lampada di calore per circa 1 ora. L&#39;ipotermia è una delle principali cause di mortalità postoperatoria e deve essere accuratamente evitata intra-e post-operatorio. Perdite di liquidi intraoperatori deve essere sostituito con un&#39;iniezione sottocutanea di 2,5 ml di soluzione salina normale, più 2,5 ml di glucosio al 5% ogni 8 ore per le prime 24 ore. I ratti dovrebbero ...

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

Orthotopic Small Bowel Transplantation in Rats

Trapianto di intestino tenue è diventata una opzione di trattamento per i pazienti con accettato irreversibile insufficienza intestinale. Il nostro modello sperimentale di trapianto ortotopico di intestino tenue nel ratto è uno strumento affidabile per affrontare immunologiche di base e processi infiammatori che complicano trapianto di intestino.

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-small-bowel-transplantation-in-rats

Research

JoVE Journal

Medicine

14.1K Views.  University of Bonn, Germany. Trapianto di intestino tenue è diventata una opzione di trattamento per i pazienti con accettato irreversibile insufficienza intestinale. Il nostro modello sperimentale di trapianto ortotopico di intestino tenue nel ratto è uno strumento affidabile per affrontare immunologiche di base e processi infiammatori che complicano trapianto di intestino.

Video: Orthotopic Small Bowel 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 ...

Ricerca

Medicina

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

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 ...

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. ...

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 ...

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis. In recent years, oncolytic virotherapies have proven themselves to be potentially powerful tools to fight malignancy. Due to the unique dual blood supply in the liver, it is possible to apply therapies locally to orthotopic liver tumors, which are predominantly fed by arterial blood flow. We have previously demonstrated that hepatic arterial delivery of oncolytic viruses results in safe and efficient transduction efficiency of multifocal HCC lesions, resulting in significant prolongation of survival in immune competent rats. This procedure closely mimics the application of transarterial embolization in patients, which is the standard palliative care provided to many HCC patients. The ability to administer tumor therapies through the hepatic artery in rats allows for a highly sophisticated preclinical model for evaluating novel viral vectors under development. Here we describe the detailed protocol for microdissection of the hepatic artery for infusion of oncolytic virus vectors to treat orthotopic HCC.

Oncolytic virotherapies are under development as novel therapeutics for the treatment of hepatocellular carcinoma (HCC). Here we describe a method for locoregional therapy of HCC via hepatic arterial administration of oncolytic virus.

Transarteriosa somministrazione di oncolitici virus per locoregionale Terapia delle ortotopico HCC in Rats

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis.  In recent years, oncolytic virotherapies have proven ...

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 ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

Trapianto di rene ortotopico ratto: un nuovo approccio chirurgico semplificato

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...

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 ...

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

Auto-trapianto di rene ortotopico in un modello suino utilizzando la conservazione degli organi 24 ore su 24 e la telemetria continua

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...

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. ...