1. Operação doador <ol><li> O rato dador devem ser mantidos em jejum durante 24 horas (acesso livre à água / solução de glicose). </li><li> Para induzir a anestesia inalatória de isoflurano, comece com 2% sobre atomizador padrão, e em seguida, reduzir para 1% após a realização de laparotomia. Realize uma pitada de igual para verificar sedação. </li><li> Raspar o abdômen e limpe com preparação da pele 3 vezes (Kodan). Em seguida, executar uma incisão mediana, após a administração subcutânea do analgésico. </li><li> Após o enxerto ser envolta em gaze embebida salina, separam-se as aderências fisiológicas entre o pâncreas e do cólon ascendente cuidadosamente com um Q-tip (sob o microscópio cirúrgico com ampliação 6x). </li><li> Ligadura e dividir os vasos cólica ileocecal e direita e meio usando 7-0 seda. Após o cólon ascendente foi espalhada para fora para o lado direito da veia mesentérica superior (SMV), os vasos de cólica ileocecal, direita e do meio pode ser identificado por ligadura e divisão usando 7-0 laços. </li><li> Retrair o estômago para cima, de modo que a totalidade SMV é endireitada para fora e exposta. Use uma pinça mosquito para a retração. O dispositivo de fixação é realizada em massa plasticina formado em forma, conforme necessário. </li><li> Ligadura e dividir as veias pancreatico-duodenais provenientes do SMV. Todas as pequenas veias pancreatico-duodenais provenientes do SMV deve ser cuidadosamente identificada, ligada com 7-0 seda e dividida antes do tecido pancreático pode ser removido a partir do enxerto. </li><li> Ligadura e dividir o tecido conjuntivo, incluindo todos os perder linfáticos e SMV entre a aorta abdominal. Com o enxerto ainda está do lado direito do abdómen, o tecido conjuntivo frouxo, incluindo todos os vasos linfáticos e SMV entre a aorta abdominal é acessível. Este tecido conjuntivo devem ser ligados utilizando 7-0 seda e dividido para evitar linforréia pós-operatória do enxerto intestinal. </li><li> Ligadura e dividir a artéria renal direita. Depois que o tecido conjuntivo é dividido, a artéria renal direita torna-se accessible e é ligada e dividida usando 7-0 seda. </li><li> Sistemicamente heparinise o rato, utilizando 200 unidades de heparina iv através da veia peniana. </li><li> Ligar as artérias marginais, e dividir o intestino delgado na junção duodeno-jejuno e no íleo terminal. </li><li> A aorta é ligada proximalmente para a origem da SMA. A veia porta é seccionado na confluência com a veia esplênica. Em seguida, o enxerto é colhida com o seu consistindo pedículo vascular da SMA com um segmento da aorta. </li></ol> 2. Procedimento Backtable <ol><li> Perfundir o enxerto com 3 ml de solução da Universidade de Wisconsin (UW), a 4 ° C, através da conduta aórtica e irrigar o lúmen intestinal a partir do final do jejuno, com 30 ml de solução Nebacetin Uro-N, a 4 ° C (a irrigação do lúmen é obrigatório) . </li></ol> Imediatamente após a extracção do enxerto, o conduto aórtico é utilizada para a perfusão, com 3 ml de solução gelada de UW. Para isto, um cateter de 20 G em ivuma seringa de 10 ml. O perfusato deve ser observada a fluir livremente para fora a partir da veia portal dividido. Para a irrigação intestinal com Nebacetin, uma seringa de 50 ml. <ol start="2"><li> Armazene a solução de enxerto de UW, a 4 ° C durante a preparação do receptor. </li></ol> 3. Operação destinatário <ol><li> O rato receptor devem ser mantidos em jejum durante 24 horas (acesso livre à água / solução de glicose). </li><li> Para induzir a anestesia inalatória de isoflurano, comece com 2% sobre atomizador padrão, e em seguida, reduzir para 1% após a realização de laparotomia. Realize uma pitada de igual para verificar sedação. </li><li> Raspar o abdômen e limpe com preparação da pele 3 vezes (Kodan). Em seguida, executar uma incisão mediana, após a administração subcutânea de carprofeno 5 mg / kg sc de analgesia. </li><li> Enrole o intestino destinatário em gaze embebida soro fisiológico e colocá-lo no peito do destinatário. </li><li> Abra o retroperitônio sem rodeios com Q-dicas, e expor a abdominal aorta ea veia cava inferior, logo abaixo dos vasos renais para baixo para o nível dos vasos ilíacos, se necessário também usar microtesoura. Ligar o tribuaries pequenas e lombar da coluna vertebral da aorta e veia cava usando 7-0 seda para evitar a perda de sangue. (Para fazer isso, altere o zoom microscópio cirúrgico até 16x.) </li><li> Atravesse-braçadeira a aorta ea veia cava inferior abaixo dos vasos renais esquerda proximal e acima da bifurcação ilíaca distalmente usando clipes de microvasos. Apenas uma braçadeira é utilizada proximal distal, bem como para fixar ambos os navios simultaneamente. Incisão ambos os navios anteriormente usando um microlanceta e lavar o sangue restante. </li><li> Criar uma anastomose término-lateral entre o segmento da aorta infra-renal da aorta do enxerto e do destinatário, usando uma sutura continua de prolene 10-0 .. Inicialmente, o enxerto é colocado no lado direito do abdómen (a cabeça do rato posicionado a 12 horas) para realizar os pontos da parede de trás da anastomose arterial e atar o fio de sutura inferior estadia. Then, o enxerto é virado para o lado esquerdo do abdómen (a cabeça do rato posicionado a 09:00), para expor e sutura da parede frontal da anastomose. </li><li> Anastomose término-lateral entre a veia porta do doador e do receptor IVC é realizada do mesmo modo, executando usando suturas 10-0 Prolene. Com o rato ainda lateralmente deitadas (cabeça na posição 9 horas), o enxerto é colocado no lado esquerdo do abdómen inferior e um fio de sutura é colocado estadia. A anastomose é iniciado com a parede do fundo do interior do vaso. Após a sutura está ligada estadia inferior, os pontos da parede frontal podem ser executadas a partir do exterior. </li><li> Remover os grampos distais primeiro lugar, seguido pelos grampos superiores. Qualquer sangramento anastomótico é controlada pela pressão direta usando Q-dicas. O enxerto deve ser verificado para igual e reperfusão rápida. </li><li> Ressecar intestino delgado do destinatário inteiro após a ligadura dos vasos mesentéricos. Destinatários enterectomizados subtotal, preservando 2-3 cm de projejuno ximal e 1 cm do íleo terminal. </li><li> Restaurar a continuidade entérico por proximal (jejuno-jejunostomia) e distal (íleo-ileostomia) de ponta a ponta anastomoses intestinais utilizando uma sutura interrompida uma camada com 6-0 Monocryl. Cerca de 16 pontos são necessários para completar as anastomoses. </li><li> Irrigar a cavidade peritoneal com solução salina normal até ficar limpo. Administrar 2 ml de soro fisiológico por via intraperitoneal para a reposição de líquidos. Em seguida, feche o abdômen com uma sutura contínua com Vicryl 3-0 para a camada muscular, mais uma sutura da pele contínua. </li><li> No pós-operatório os ratos devem ser mantidos em jejum (com acesso a água e uma solução de glucose) para um outro 24 hr depois reiniciado em dieta padrão e água ad libitum. Analgesia com carprofeno deve ser administrada durante 3 dias (ver dosagem abaixo). </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>

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Embora os modelos de transplante de intestino de ratos têm sido descritos, o mais cedo no 5 1970ies a maioria dos modelos recentemente utilizados envolvem transplante heterotópico intestinal utilizando diferentes técnicas 13. Enquanto os procedimentos heterotópico em geral, têm a vantagem de técnicas de microcirurgia mais fáceis e mais fácil acessibilidade do enxerto para a avaliação, o transplante heterotópico intestinal tem a grande desvantagem de não levar em conta as múltiplas inte...

<table border="1"><tbody><tr><td> Nome do reagente </td><td> Companhia </td><td> Número de catálogo </td><td> Comentários (opcional) </td></tr><tr><td> Universidade de Wisconsin (UW) solução (ViaSpan) </td><td> Bristol-Myers Squibb </td><td></td><td></td></tr><tr><td> Uro-Nebacetin solução N </td><td> Nycomed </td><td> 6967855 </td><td></td></tr><tr><td> Ampicilina </td><td> Ratiopharm </td><td></td><td></td></tr><tr><td> Carprofeno (Rimadyl) </td><td> Pfizer </td><td></td><td></td></tr><tr><td> Prolene sutura 10-0 unresorbable </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Monocryl sutura reabsorvível 6-0 </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Fio de vicryl 3-0 reabsorvível </td><td> Ethicon </td><td></td><td></td></tr><tr><td> Cateter iv G 20 milímetros 1.1x33 </td><td> Braun </td><td></td><td></td></tr><tr><td> Cateter iv G 22 0.9x25 milímetro </td><td> Braun </td><td></td><td></td></tr><tr><td> Kodan pele Prep </td><td> Schülke </td><td></td><td></td></tr><tr><td> Solução de NaCl a 0,9% Infusão </td><td> Braun </td><td></td><td></td></tr><tr><td> Fórceps curvos pequena </td><td> FineScienceTools </td><td> 11009-13 </td><td></td></tr><tr><td> Fórceps micro curvo </td><td> AESCULAP </td><td> BD 333 </td><td></td></tr><tr><td> Fórceps micro curvo </td><td> AESCULAP </td><td> FD281R </td><td></td></tr><tr><td> Micro pinça reta 1 </td><td> WPI </td><td> 5 </td><td></td></tr><tr><td> Micro pinça reta 2 </td><td> WPI </td><td> 2 </td><td></td></tr><tr><td> Suporte da agulha micro </td><td> WPI </td> &lt;td&gt; 14081 </td><td></td></tr><tr><td> Micro tesoura </td><td> FineScienceTools </td><td> 15006-09 </td><td></td></tr><tr><td> Micro bisturi </td><td> MANI </td><td> Faca oftálmica </td><td></td></tr><tr><td> Grampos Micro </td><td> AESCULAP </td><td> FB329R </td><td></td></tr></tbody></table>

orthotopic small bowel transplantation in rats

Transplante de intestino delgado, tornou-se uma opção clínico aceite para doentes com síndrome do intestino curto e insuficiência de nutrição parentérica (insuficiência intestinal irreversível). Em centros especializados melhorou operatório e estratégias de gestão levaram a paciente a curto e médio prazo e excelente sobrevida do enxerto, proporcionando alta qualidade de vida 1,3. Ao contrário do transplante mais comum de outros órgãos sólidos (coração, fígado ou seja,) muitos mecanismos subjacentes da função do enxerto e alterações imunológicas induzidas por transplante intestinal não são totalmente conhecidos 6,7. Episódios de rejeição aguda, septicemia e insuficiência crônica do enxerto são os principais obstáculos ainda contribuem para menos favorável resultado a longo prazo e dificultando um emprego mais amplo do procedimento, apesar de um número crescente de pacientes em nutrição parenteral domiciliar que potencialmente podem beneficiar de um transplante de tal. O intestino delgado contémum grande número de leucócitos passageiros comumente referida como parte do intestino associada sistema linfóide (GALT), esta sendo parte da razão para a alta immunogenity do enxerto intestinal. A presença ea proximidade de muitos comensais e patogénicos no intestino explica a gravidade dos episódios de sépsis, uma vez a integridade da mucosa do enxerto for comprometida (por exemplo, rejeição). Para avançar no campo de dados de múltiplos órgãos intestinais e transplante mais gerados a partir de modelos animais confiáveis ​​e viáveis ​​é necessário. O modelo aqui fornecida combina confiabilidade e viabilidade uma vez estabelecido de forma padronizada e pode fornecer informações valiosas nas subjacentes complexos moleculares, os mecanismos celulares e funcionais que são acionados por transplante intestinal. Nós temos utilizado com sucesso e refinado o procedimento descrito ao longo de mais de 5 anos no nosso laboratório 8-11. O formato de vídeo baseado JoVE é especialmente útil para demonstrar o procedimento complexoDure e evitar as armadilhas iniciais para grupos de planejamento para estabelecer um modelo de roedor ortotópico investigando o transplante intestinal.

Evolução pós-operatória normal Os animais transplantados deve se recuperar rapidamente do procedimento sob uma lâmpada de calor por cerca de 1 hora. A hipotermia é uma das principais causas de mortalidade no pós-operatório e devem ser evitados intra e pós-operatório. As perdas de fluido intra-operatória deve ser substituído por injeção subcutânea de 2,5 ml de solução salina normal mais 2,5 ml de glicose 5% a cada 8 horas para a primeira hora 24. Os ratos também devem ter livre...

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

Orthotopic Small Bowel Transplantation in Rats

Transplante de intestino delgado tornou-se uma opção de tratamento aceitável para os pacientes com falência intestinal irreversível. Nosso modelo experimental de transplante intestinal ortotópico pequena em ratos serve como uma ferramenta confiável para enfrentar imunológico subjacente e os processos inflamatórios que complicam o transplante intestinal.

Transplante de intestino delgado de ratos ortotópico

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 visualizações.  University of Bonn, Germany. Transplante de intestino delgado tornou-se uma opção de tratamento aceitável para os pacientes com falência intestinal irreversível. Nosso modelo experimental de transplante intestinal ortotópico pequena em ratos serve como uma ferramenta confiável para enfrentar imunológico subjacente e os processos inflamatórios que complicam o transplante intestinal.

Vídeo: 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.

Desenvolvimento de bronquiolite obliterante em um modelo murino de transplante de pulmão ortotópico

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

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.

Transplante hepático ortotópico experimental em ratos

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.

Transplante ortotópico de aorta em camundongos para o Estudo da Doença Vascular

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).

Ortotópico Hind Limb Transplantation no 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.

Transarterial Administração de Oncolíticos Vírus para loco-regional Terapia de Ortotópico HCC em Ratos

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.

Um modelo pré-clínico porcino de transplante cardíaco ortotópico

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.

Transplante de rim de rato ortotópico: uma abordagem cirúrgica nova e simplificada

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.

Complicações reduzidas após reconexão arterial em um modelo de rato de transplante de fígado ortotópico

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-transplante ortotópico de rim em um modelo suíno usando preservação de órgãos 24 horas e telemetria contínua

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.

Transplante ortotópico de pulmão esquerdo em modelo suíno juvenil para PESL

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