The study was supported by research grants from the Roche Organ Transplant Research Foundation (ROTRF) and Astellas. Markus Selzner was supported by an ASTS Career Development Award. Matthias Knaak was supported by the Astellas Research Scholarship. We thank Uwe Mummenhoff and the Birmingham family for their generous support.

<ol>
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Sci. 19 (2), 131-136 (1989).</li><li>Terblanche, J. <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+homotransplantation:+an+experimental+study+in+the+unmodified+pig.">Orthotopic liver homotransplantation: an experimental study in the unmodified pig.</a> S. Afr. Med. J. 42 (20), 486-497 (1968).</li><li>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=Prolonged+survival+of+liver+transplants+in+the+pig.">Prolonged survival of liver transplants in the pig.</a> Br. Med. J. 4 (5580), 645-648 (1967).</li><li>Battersby, C., Egerton, W. S., Balderson, G., Kerr, J. 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V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermic+machine+perfusion+of+liver+grafts+for+transplantation:+technical+development+in+human+discard+and+miniature+swine+models.">Hypothermic machine perfusion of liver grafts for transplantation: technical development in human discard and miniature swine models.</a> Transplant Proc. 37 (1), 323-325 (2005).</li><li>Minor, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermic+reconditioning+by+gaseous+oxygen+improves+survival+after+liver+transplantation+in+the+pig.">Hypothermic reconditioning by gaseous oxygen improves survival after liver transplantation in the pig.</a> Am. J. Transplant. 11 (12), 2627-2634 (2011).</li><li>Kelly, D. 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The authors have nothing to disclose.

Sperimentale suina OLTx è una procedura difficile per un ambiente di ricerca senza le risorse di terapia intensiva di uno scenario clinico. Le possibili complicanze comprendono instabilità emodinamica, emorragia, organo ischemia, ipotermia, e metaboliche, così come le vie respiratorie, scompenso. Per ogni gruppo di ricerca, la formazione procedurale sufficiente della tecnica chirurgica 5 nonché al 14,15 anestesia maiale è obbligatoria per ottenere risultati rappresentativi e riproducibili. Molti sottigliezze tecniche sono state descritte in letteratura, in particolare per quanto riguarda la fase di ricostruzione vascolare 5. Il protocollo OLTx sopra descritto fornisce le informazioni necessarie per un modello-cava sostituzione assomiglia OLTx umano. I risultati forniti dimostrano la sopravvivenza animale affidabile e di recupero del trapianto in entrambi i modelli HBD e DCD. Il protocollo è applicabile in situazioni di sopravvivenza a breve termine utilizzati nel trapianto riperfusione esperiments, per esempio, così come nei modelli di sopravvivenza a lungo termine, come studi di tolleranza. Un grande ostacolo della suina OLTx è relativamente scarsa tolleranza di cava e vena porta clampaggio. Congestione splancnica durante la fase anepatica provoca ipertensione venosa e capillare danni che possono portare a importanti ischemia intestinale e instabilità emodinamica al punto di uno shock irreversibile anche dopo organo riperfusione 7. Poiché la vena cava è completamente annegato nel parenchima epatico, una procedura piggy-back-cava conservazione non è fattibile. L&#39;occlusione totale della vena cava durante la fase di ricostruzione cava compromette la stabilità emodinamica del maiale. Anche se alcuni rapporti indicano che suina OLTx può essere eseguita durante totale cava e l&#39;occlusione della vena porta inferiore a 25 min 16,17, una tecnica di bypass porto-cava-giugulare per il tempo della ricostruzione vascolare è l&#39;opzione più sicura e più pratico 7- 9,18. In thesperienza e degli autori, un passivo porto-giugulare bypass non è ottimale per mantenere il emodinamicamente maiale stabile durante la fase anepatica. Il modello di bypass, compresa la decompressione attiva della cava sottoepatica e vena porta, permette una fase di ricostruzione tranquillità delle anastomosi cavali e portale sovraepatiche anche con il tempo di bloccaggio prolungato a causa di complicazioni impreviste. Contrariamente alle precedenti relazioni 7, una splenectomia non è obbligatoria quando il catetere portale di bypass viene rimosso. Sia splenica e la vena sono chiusi a metà circa la lunghezza della milza lasciando la metà prossimale sufficientemente perfusione. Complicazioni come sanguinamento o embolia gassosa a causa di bypass disconnessione sono evitabili, garantendo che il bypass è posizionato accuratamente e fissato correttamente. In sopravvivenza OLTx esperimenti a lungo termine, l&#39;anastomosi dotto biliare è considerato un punto debole a causa del suo alto tasso di complicanze 19. Il tessuto biliare è molto fragile e needs particolare attenzione quando viene gestita. Molte tecniche di anastomosi diverse sono state descritte 5,19. Una anastomosi end-to-end è tecnicamente facile e associata a complicazioni minime 19. Una sutura continuo con un ago non tagliente comprese grandi sezioni di tessuto connettivo peribiliare sembra essere superiore ad una sutura interrotta. Il dotto biliare è posto sotto tensione inutile quando i singoli punti di sutura interrotta sono annodati. Ciò può causare rotture del tessuto e perdite biliari consecutivi. Il materiale di sutura - riassorbibili o non riassorbibili - di solito non è importante, data la sua durata limitata fino a quando il maiale è terminata. Per i modelli di sopravvivenza a lungo termine nell&#39;arco di diversi mesi, suture assorbibili - come in OLTx umano - sono preferibili. Particolare attenzione deve essere presa con il post-operatorio di follow-up. Nutrizione sufficiente e alimentazione del fluido, un protocollo di sollievo dal dolore affidabile, e una adeguata immunosoppressioneregime sono obbligatori. Per gli esperimenti a lungo termine, l&#39;immunosoppressione appare particolarmente importante. Rispetto ad altri mammiferi, i maiali mostrano sorprendentemente basso tasso di rigetto immunologico dopo OLTx 20,21. Infiltrazioni di cellule rotonde sono la massima durante la seconda settimana dopo il trapianto e diminuiscono spontaneamente anche senza immunosoppressione. Il rifiuto è raramente la causa di morte dopo suina OLTx 22. Tuttavia, anche con il protocollo di immunosoppressione coinvolge la somministrazione di steroidi iv e inibitori della calcineurina po accennato qui, il rigetto del trapianto è indicato da un lieve aumento delle transaminasi a partire da circa 4 giorni dopo OLTx e confermato dal campo apparente portale infiltrazione di cellule turno. Gli inibitori della calcineurina può essere somministrato sia po 23,24 o IV 25,26; Entrambi i metodi hanno svantaggi. Anche con ausili applicativi orali, l&#39;importo effettivo raggiungimento del tratto gastro-intestinale resta sfuggente. D&#39;altra parte, iv infusione continua inpenna di un maiale con un animale attivo è difficile. Pertanto, l&#39;applicazione iv deve essere effettuata come un bolo, che si traduce in picchi di concentrazione elevata farmaco con potenziali effetti tossici. Tuttavia, entrambi i metodi di applicazione sembrano permettere la sopravvivenza a lungo termine. Simile a un contesto clinico, si raccomanda la profilassi post-operatorio di stress ulcera. Sanguinamento post-operatorio da ulcere peptiche è un problema frequente e può essere correlata a una funzione epatica 27. Dopo un paio di casi di sanguinamento gastrointestinale in entrambi i gruppi OLTx, gli autori hanno iniziato la profilassi regolari con pantoprazolo e non hanno avuto alcun sanguinamento gastrointestinale da allora. Rigoroso mantenimento delle condizioni di sterilità intraoperatoriamente, paragonabili a quelle previste in una sala operatoria clinica, e conseguenti profilassi antibiotica, diminuisce il rischio di complicanze infettive. In conclusione, questo articolo provides informazioni pratiche per la creazione di un programma di suino OLTx in un ambiente di ricerca. Sufficiente dedizione, pratica, e lavoro di squadra è importante al fine di diminuire il periodo di apprendimento, per produrre risultati affidabili, e per ridurre i costi e il numero di animali di ricerca.

Trapianto di fegato (OLTx) è l&#39;unica opzione di trattamento per i pazienti con malattia epatica allo stadio terminale o carcinoma epatocellulare avanzato. Negli ultimi 25 anni, il numero dei candidati in lista d&#39;attesa è progressivamente aumentato e ora supera di gran lunga il numero di innesti disponibili. Nella maggior parte delle regioni di trapianto, dal 20 al 30% dei pazienti in lista di attesa per trapianto di fegato muoiono senza ricevere un trapianto d&#39;organo o di esclusione dalle negoziazioni a causa della progressione della malattia. Strategie per aumentare il pool di donatori e, quindi, il numero di innesti a disposizione, sono disperatamente necessari. Criteri di assegnazione degli organi estese, conservazione trapianto prolungato, e induzione della tolleranza immunologica rappresentano ancora grandi sfide cliniche 1-3. Quindi, la ricerca sperimentale OLTx è fondamentale al fine di ottimizzare clinica pratica OLTx. Suini OLTx è un modello sperimentale consolidata che assomiglia OLTx umano in molti modi, tra cui ldimensioni Iver, l&#39;anatomia, la fisiologia e 4-6. Così, è diventato un metodo sperimentale standard campi di ricerca quali tecniche chirurgiche, la fisiologia, l&#39;immunologia, la conservazione, e danno da ischemia-riperfusione. Numerose tecniche di approvvigionamento graft, destinatario epatectomia, e, soprattutto, la ricostruzione vascolare, sono stati descritti in letteratura 5. La scelta della tecnica appropriata varia in base alle preferenze del ricercatore e capacità tecnica. Contrariamente allo scenario umano, congestione splancnico durante la fase anepatica rappresenta un problema importante in porcine OLTx. La successiva ischemia intestinale e danno vascolare congestizia possono causare grave instabilità emodinamica, mettendo a repentaglio la sopravvivenza di maiale e, quindi, il successo dell&#39;esperimento 7-9. Pertanto, sufficiente la decompressione intestinale è obbligatorio, soprattutto in contesti sperimentali tecnicamente meno raffinati. Noiing un porto-cavale-giugulare shunt attivo per tutta la durata della fase di anepatica è un&#39;opzione affidabile per evitare la congestione intestinale. Il sistema può essere utilizzato per esperimenti riperfusione precoce nonché scenari di sopravvivenza a lungo termine. Il protocollo che segue contiene tutte le informazioni per un fegato modello di trapianto di stabili e riproducibili nei suini, compresi gli appalti di fegato da donatore, operazione destinatario, incluso epatectomia e tecniche di ricostruzione nave end-to-end, e cure post-operatorie.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Atropine Sulphate 15mg/30mL</td>
			<td style="width:257px;">Rafter 8 Products</td>
			<td align="right" style="width:201px;">238481</td>
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			<td height="20" style="height:20px;width:276px;">Buprenorphine 0.3mg/mL</td>
			<td style="width:257px;">RB Pharmaceuticals LDT</td>
			<td style="width:201px;">N/A</td>
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			<td height="19" style="height:19px;width:276px;">Cefazolin 1g</td>
			<td style="width:257px;">Pharmaceutical Partners of Canada Inc.</td>
			<td align="right" style="width:201px;">2237138</td>
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			<td height="20" style="height:20px;width:276px;">Cyclosporin Oral Solution 5000mg/50mL</td>
			<td style="width:257px;">Novartis Pharmaceuticals Canada Inc.</td>
			<td align="right" style="width:201px;">2150697</td>
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			<td height="19" style="height:19px;width:276px;">Fentanyl Citrate 0.25mg/5mL</td>
			<td style="width:257px;">Sandoz Canada Inc.</td>
			<td align="right" style="width:201px;">2240434</td>
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			<td height="19" style="height:19px;width:276px;">Heparin 10,000iU/10mL</td>
			<td style="width:257px;">Leo Pharma A/S</td>
			<td align="right" style="width:201px;">453811</td>
			<td style="width:240px;"></td>
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			<td height="19" style="height:19px;width:276px;">Isoflurane 99.9%, 250mL</td>
			<td style="width:257px;">Pharmaceutical Partners of Canada Inc.</td>
			<td align="right" style="width:201px;">2231929</td>
			<td style="width:240px;"></td>
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			<td height="20" style="height:20px;width:276px;">Ketamine Hydrochloride&#160; 5000mg/50mL</td>
			<td style="width:257px;">Bimeda-MTC Animal Health Inc.</td>
			<td align="right" style="width:201px;">612316</td>
			<td style="width:240px;"></td>
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			<td height="20" style="height:20px;width:276px;">Lactated Ringer&#8217;s + 5% Dextrose, 0.5L</td>
			<td style="width:257px;">Baxter Corporation</td>
			<td align="right" style="width:201px;">61131</td>
			<td style="width:240px;"></td>
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			<td height="20" style="height:20px;width:276px;">Lacteted Ringer&#8217;s, 1L</td>
			<td style="width:257px;">Baxter Corporation</td>
			<td align="right" style="width:201px;">61085</td>
			<td style="width:240px;"></td>
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			<td height="20" style="height:20px;width:276px;">Metronidazole 500mg/100mL</td>
			<td style="width:257px;">Baxter Corporation</td>
			<td align="right" style="width:201px;">870420</td>
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			<td height="20" style="height:20px;width:276px;">Midazolame 50mg/10mL</td>
			<td style="width:257px;">Pharmaceutical Partners of Canada Inc.</td>
			<td align="right" style="width:201px;">2242905</td>
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			<td height="20" style="height:20px;width:276px;">Pantoprazole 40mg</td>
			<td style="width:257px;">Sandoz Canada Inc.</td>
			<td align="right" style="width:201px;">2306727</td>
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			<td height="20" style="height:20px;width:276px;">Potassium Chloride 40mEq/20mL</td>
			<td style="width:257px;">Hospira Healthcare Corporation</td>
			<td align="right" style="width:201px;">37869</td>
			<td style="width:240px;"></td>
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			<td height="20" style="height:20px;width:276px;">Propofol 1000mg/100mL</td>
			<td style="width:257px;">Pharmascience Inc.</td>
			<td align="right" style="width:201px;">2244379</td>
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		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Protamine Sulfate 50mg/5mL</td>
			<td style="width:257px;">Pharmaceutical Partners of Canada Inc.</td>
			<td align="right" style="width:201px;">2139537</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Saline 0.9%, 1L</td>
			<td style="width:257px;">Baxter Corporation</td>
			<td align="right" style="width:201px;">60208</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Sodium Bicarbonate 50 mEq/50mL</td>
			<td style="width:257px;">Hospira Healthcare Corporation</td>
			<td align="right" style="width:201px;">261998</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Solu-Medrol 500mg</td>
			<td style="width:257px;">Pfizer Canada Inc.</td>
			<td align="right" style="width:201px;">2367963</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Tranexamic Acid 1000mg/10mL</td>
			<td style="width:257px;">Pfizer Canada Inc.</td>
			<td align="right" style="width:201px;">2064413</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">University of Wisconsin Solution, SPS-1</td>
			<td style="width:257px;">Organ Recovery Systms</td>
			<td style="width:201px;">SPS-1</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Xylocaine Endotracheal 10mg/50mL</td>
			<td style="width:257px;">AstraZeneca</td>
			<td align="right" style="width:201px;">2003767</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Appose ULC 35W skin stapler</td>
			<td style="width:257px;">Covidien Canada</td>
			<td align="right" style="width:201px;">803712</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Maxon, 1</td>
			<td style="width:257px;">Covidien Canada</td>
			<td align="right" style="width:201px;">606173</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Sofsilk, 0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">S606</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Sofsilk, 2-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">S405</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Sofsilk, 3-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">S404</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 4-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP581X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 5-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP725X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Surgipro II, 6-0</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">VP733X</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;"></td>
			<td style="width:257px;"></td>
			<td style="width:201px;"></td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Catheter i.v, 18 G</td>
			<td style="width:257px;">BD Canada</td>
			<td align="right" style="width:201px;">381147</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Cook TPN catheter, 9.5Fr</td>
			<td style="width:257px;">Cook Medical Company</td>
			<td style="width:201px;">C-TPNS-9.5-90</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">PSI Kit for sheath catheter, 8.5Fr</td>
			<td style="width:257px;">Arrow International</td>
			<td style="width:201px;">ASK-09803-UHN</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Infusion Pump Line</td>
			<td style="width:257px;">Smith Medical ASD Inc.</td>
			<td style="width:201px;">21-0442-25</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Liver Admin Set (flush line)</td>
			<td style="width:257px;">CardioMed Supplies Inc</td>
			<td align="right" style="width:201px;">17175</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Mallinckrodt, Tracheal Tube, 6.5mm</td>
			<td style="width:257px;">Covidien Canada</td>
			<td align="right" style="width:201px;">86449</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Med-Rx Suction Connecting Tube</td>
			<td style="width:257px;">Benlan Inc.</td>
			<td style="width:201px;">70-8120</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Organ Bag</td>
			<td style="width:257px;">CardioMed Supplies Inc</td>
			<td align="right" style="width:201px;">2990</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Suction Tip</td>
			<td style="width:257px;">Tyco Healthcare Group LP</td>
			<td align="right" style="width:201px;">8888501023</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Valleylab, Cautery Pencil</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">E2515H</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Valleylab, Patient Return Electrode</td>
			<td style="width:257px;">Covidien Canada</td>
			<td style="width:201px;">E7507</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;"></td>
			<td style="width:257px;"></td>
			<td style="width:201px;"></td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector 3/8&#8221; x 1/4&#8220;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">955083-001</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector 3/8&#8221; x 3/8&#8221; Luer Lock</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">955163-001</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Connector Y 3/8&#8221; x 3/8&#8221; x 1/4&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">961360-002</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Tubing 1/4&#8221; x 1/16&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">039505-010</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Bypass Tubing 3/8&#8221; x 3/32&#8221;</td>
			<td style="width:257px;">Raumedic AG</td>
			<td style="width:201px;">039535-005</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Rotaflow Cenrtifugal Pump</td>
			<td style="width:257px;">Maquet-Dynamed</td>
			<td style="width:201px;">HC 2821</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Stainless Steel Hose Clamp Ring, 5mm</td>
			<td style="width:257px;">Oetiker</td>
			<td align="right" style="width:201px;">16700007</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Abdominal Retractor</td>
			<td style="width:257px;">Medite GmbH</td>
			<td style="width:201px;">N/A</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">De Bakey &#8211; Beck, Infrahepatic Cava Clamp</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB519R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Diethrich, Atraumaitc Clamp (Portal Vein)</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB525R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Gregory Bull Dog Clamp, curved</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB382R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Gregory Bull Dog Clamp, straight</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB381R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Potts &#8211; De Martel, Scissors</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">BC648R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Satinsky, Suprahepatic Cava Clamp</td>
			<td style="width:257px;">Aesculap Inc.</td>
			<td style="width:201px;">FB605R</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:276px;">Symetrical Tubing Clamp</td>
			<td style="width:257px;">Codman Instruments</td>
			<td align="right" style="width:201px;">198010</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Anesthesia Machine, Optimax</td>
			<td style="width:257px;">Moduflex Anesthesia Equipment</td>
			<td style="width:201px;">SN5180</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Bypass Flow meter, HT 110</td>
			<td style="width:257px;">Transonic Systems Inc.</td>
			<td style="width:201px;">HT110B11106</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Flow meter probe, H6XL</td>
			<td style="width:257px;">Transonic Systems Inc.</td>
			<td style="width:201px;">H6Xl689</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Heat Therapy Pump, T/Pump</td>
			<td style="width:257px;">Gaymar Industries Inc</td>
			<td style="width:201px;">TP500-G89D19</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Infusion Pump 3000</td>
			<td style="width:257px;">SIMS Graseby LTD.</td>
			<td style="width:201px;">SN300050447</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Isoflurane Vapor 19.1</td>
			<td style="width:257px;">Draeger Medical Canada Inc.</td>
			<td style="width:201px;">N/A</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Monitor, Datex AS 3</td>
			<td style="width:257px;">Instrumentarium Corp./ Hitachi</td>
			<td style="width:201px;">D-VHC14-23-02</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Rotaflow Centrifugal Drive Unit</td>
			<td style="width:257px;">Marquet-Dynamed</td>
			<td align="right" style="width:201px;">952301</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Rotaflow Console</td>
			<td style="width:257px;">Marquet-Dynamed</td>
			<td align="right" style="width:201px;">706035</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Temperature Therapy Pad</td>
			<td style="width:257px;">Gaymar Industries Inc</td>
			<td style="width:201px;">TP26E</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Valleylab Force Tx</td>
			<td style="width:257px;">Valleylab Inc.</td>
			<td align="right" style="width:201px;">216151480</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Ventilator, AV 800</td>
			<td style="width:257px;">DRE Medical Equipment</td>
			<td style="width:201px;">40800AVV</td>
			<td style="width:240px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:276px;">Warm Touch, Patient Warming System</td>
			<td style="width:257px;">Nellcor/ Covidien Canada</td>
			<td style="width:201px;">5015300A</td>
			<td style="width:240px;"></td>
		</tr>
	</tbody>
</table>

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.

In un primo studio di trapianto, un modello donatore a cuore battente (HBD, n = 5) è stato confrontato con un modello DCD (n = 10) esposta a 45 min di ischemia calda in situ. In entrambi i gruppi, gli innesti sono stati conservati in ghiaccio per 10 ore dopo l&#39;approvvigionamento. Nel gruppo HBD, il 100% dei suini beneficiari sopravvisse fino alla fine del follow-up il giorno 5 dopo il trapianto. Nel gruppo DCD, solo il 50% dei suini beneficiari sopravvissuto per 5 giorni a causa di problemi di coagulazione o scompenso metabolico, come risultato della funzione epatica post-operatorio diminuito. Tutti i campioni di sangue sono stati prelevati dal catetere venoso centrale. Dopo la centrifugazione, campioni di siero sono stati ottenuti e analizzati per danno epatocellulare (aspartato aminotransferasi, AST), la funzione delle vie biliari (bilirubina totale e fosfatasi alcalina), e la funzione del fegato (INR). Il tempo impiegato per ogni marcatore è illustrata nelle figure 2 - 5. <p class="jove_content"&gt; AST livelli hanno raggiunto un picco dopo 24 ore (1414 ± 538 U / L nel gruppo HBD e 2.296 ± 1.313 U / L nel gruppo DCD, p = 0,13) e sono tornati a valori quasi normali dopo 5 giorni. Analogamente i valori della fosfatasi alcalina sono stati notevolmente aumentati dopo 36 ore nel gruppo DCD (224 ± 111 U / L) rispetto al gruppo HBD (162 ± 54 U / L, p = 0,27). Mentre bilirubina totale è rimasta stabile nel gruppo HBD (≤ 10 micromol / L per tutto), è progressivamente aumentato nel gruppo DCD fino al giorno 5 (23 ± 31 micromol / L, p = 0,43). La grande deviazione standard dei valori di bilirubina nel gruppo DCD mostra una peculiare eterogeneità infortunio biliare in questo gruppo. INR come marker della funzione epatica ha mostrato un andamento simile ai valori AST. Valori ha raggiunto un picco a 24 ore e sono stati ripristinati i valori quasi normali dopo 5 giorni. Il gruppo HBD aveva valori inferiori con un picco a 1,47 ± 0,34 rispetto al gruppo DCD (picco 1,70 ± 0,36, p = 0,32). Figura 1. Schema dello shunt porto-cavale-giugulare. Il bypass è riempita con una soluzione di Ringer lattato. Poi la parte cava viene serrato con un morsetto del tubo, le parti giugulari e splenici sono collegati ai cateteri pre-impostati, il bypass è aperto, e la pompa centrifuga viene avviato dopo portale serraggio venosa. Dopo resezione epatica, la parte cavale della tangenziale viene inserito e fissato nella infraepatica moncone vena cava, craniale alle vene renali. Il morsetto del tubo viene rilasciato per consentire la decompressione cava, oltre alla decompressione portale. <img alt="Figura 2" src="/files/ftp_upload/52055/52055fig2.jpg" /> Figura 2. aspartato aminotransferasi (AST) (HBD n = 5, DCD n = 10). AST è un marcatore sensibile di danno epatocellulare. La poppa di piccoer 24 ore è più bassa nella HBD rispetto al gruppo DCD, suggerendo il danno da riperfusione meno epatica; la deviazione standard più piccola mostra risultati più omogenei nel gruppo HBD. <img alt="Figura 3" src="/files/ftp_upload/52055/52055fig3.jpg" /> Figura 3. bilirubina totale (HBD n = 5, DCD n = 10). Bilirubina totale, come un marcatore della clearance biliare e l&#39;integrità del dotto biliare, mostra un andamento stabile ed omogenea con valori inferiori a 10 mmol / L nel gruppo HBD. La curva bilirubina nel gruppo DCD aumenta gradualmente nel tempo e mostra una deviazione standard alto, suggerendo lesioni biliare in solo una parte del gruppo sperimentale. <img alt="Figura 4" src="/files/ftp_upload/52055/52055fig4.jpg" /> Figura 4. Fosfatasi alcalina (HBD n = 5, DCD n = 10). Fosfatasi alcalina is un indicatore di danno biliare. I valori per il gruppo HBD sono inferiori rispetto a quella del gruppo DCD, che implica meno lesioni biliare. <img alt="Figura 5" src="/files/ftp_upload/52055/52055fig5.jpg" /> Figura 5. INR (HBD n = 5, DCD n = 10). Un alto valore di INR indica diminuita funzione epatocellulare a causa della diminuzione rilascio dei fattori della coagulazione. In entrambi i gruppi HBD e DCD, i valori INR tornare a valori normali 5 giorni dopo il trapianto, suggerendo il recupero della funzionalità epatica. I valori per il gruppo HBD appaiono inferiori.

Watch this Scientific Journal Video about Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt at JoVE.com

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

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 ...

technique-porcine-liver-procurement-orthotopic-transplantation-using

Nanyang Technological University

Research

JoVE Journal

Medicine

16.3K Views.  Toronto General Hospital. 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.

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

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 ...

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 ...

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 ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

Utilizzando quantitativa real-time PCR per la determinazione cellulare attecchimento dei donatori in un modello murino competitivo trapianto di midollo osseo

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

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 ...

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.

Utilizzando la tecnica di manica in un modello murino di trapianto aortico - un Video didattico

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

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 ...

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 ...