Name: heterotopic cervical heart transplantation in mice
Uploaded: 2015-08-25T14:00:00.000+00:00
Duration: 9 min 48 s
Description: Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

M.-A. Deutsch is supported by Dr. Rusche Forschungsprojekt (2014) Deutsche Stiftung f&#252;r Herzforschung and Deutsche Gesellschaft f&#252;r Thorax- Herz- und Gef&#228;&#223;chirurgie. M. Krane is supported by Deutsche Stiftung f&#252;r Herzforschung (F/37/11), Deutsches Zentrum f&#252;r Herz Kreislauf Forschung (DZHK B 13-050A; DZHK B 14-013SE) and Deutsche Forschungsgemeinschaft (KR3770/7-1; KR3770/9-1)

Todos os experimentos foram aprovados pelo alemão Heart Center Munique eo Governo da Baviera, Alemanha (Gz. 55.2-1-54-2532.3-17-13). Os animais foram alojados em uma instalação livre de patógenos específicos e recebeu cuidados humanos em conformidade com os &quot;Princípios de Laboratório Animal Care&quot; formuladas pela Sociedade Nacional de Pesquisa Médica e do &quot;Guia para o Cuidado e Uso de Animais de Laboratório&quot; fornecido pela Academia Nacional de Ciências e publicado pelos Institutos Nacionais de Saúde (NIH publicação 86-23 no., revista, 1985). NOTA: Um conjunto completo de instrumentos de microcirurgia estéreis, incluindo duas pinças curvas e dilatadores dos vasos, é necessário para o procedimento (Figura 2a). Profilaxia com antibióticos perioperatórios não é rotineiramente necessária, mas pode ser indicada em animais imunocomprometidos. Os animais foram sacrificados usando inalação de isoflurano terminal. Doador de Preparação e Graft Colheita &lt;ol&gt; <li> Anestesiar o animal por via intraperitoneal (ip) de injecção de midazolam (5,0 mg / kg), medetomidina (0,5 mg / kg) e fentanil (0,05 mg / kg de peso corporal). Posicione o mouse em decúbito dorsal sobre o campo operatório e confirmar tolerância cirúrgico pelo reflexo interdigital. </li><li> Desinfectar a pele três vezes com clorexidina e realizar uma incisão na linha média abdominal. Recolha os intestinos com gaze húmida para a esquerda para expor a veia cava inferior. </li><li> Injectar 50 UI de heparina diluído em 0,4 ml de solução salina através da veia cava inferior para a anticoagulação. Faça uma incisão na embarcação 1 min após heparinização. </li><li> Abra o diafragma e executar uma toracotomia bilateral para expor o coração. A parede anterior do tórax é refletido superiormente e mantido no lugar com um grampo. </li><li> Remova o timo e sem rodeios dissecar o tecido conjuntivo entre aorta e tronco pulmonar. Faça uma incisão na veia cava superior direita. </li><li> Perfurar a aorta ascendente mobilizado no nível doTBE com uma agulha G 30, montado sobre uma seringa de 5 ml e perfundir lentamente o coração retrogradually sem pressão excessiva, com 3 ml de 4 ° C, solução cardioplégica a frio. Evitar a entrada de ar para dentro da circulação coronária. Largar solução salina gelada para o enxerto para aumentar ainda mais a parada cardíaca. </li><li> Transecto a aorta proximal à origem do tronco braquiocefálico. </li><li> Ligadura da veia inferior e ambos cavas superior perto do coração com 8-0 seda e transecto-los para distal para as ligaduras. </li><li> Dissecar livre do tronco pulmonar como distalmente quanto possível. Ligadura das artérias pulmonares perto da bifurcação com 8-0 seda e transecto-los para distal. </li><li> Ligadura das veias pulmonares todos juntos com um único ligadura 8-0 seda e transecto-los. </li><li> Tiraram o coração do doador e armazená-lo em solução cardioplégica gelada até a implantação. </li></ol> 2. Destinatário Preparação <ol><li> Prepararos punhos para anastomoses por corte do tubo de poliimida com um bisturi n ° 11 ou uma tesoura ao microscópio. A braçadeira é constituída por um corpo cilíndrico e uma extensão (1/3 da circunferência) para a colocação da pinça vascular, cada uma com um comprimento de 1 mm (Figura 2b). </li><li> Anestesiar o animal e confirmar tolerância cirúrgico, tal como descrito acima. Injectar 0,25 ml de soro fisiológico morno solução por via subcutânea (sc) para a reposição de líquidos. </li><li> Remover os pêlos da região cervical direita usando um barbeador elétrico e colocar o animal em decúbito dorsal sobre uma almofada quente. Prenda os pés com fios de fita, evitando overstretching os membros dianteiros, pois isso pode comprometer a respiração. Cubra os olhos usando pomada oftálmica para prevenir o ressecamento. Desinfetar o local da cirurgia três vezes com clorexidina. </li><li> Faça uma incisão transversa da pele a partir do ângulo da mandíbula direito ao entalhe jugular. </li><li> Sem rodeios mobilizar a extErnal veia jugular. Cauterizar ramos laterais com um fórceps bipolares e transecto-los. </li><li> Dividir a veia jugular externa entre ligaduras no nível de confluência com a veia a partir da glândula submandibular. </li><li> Puxe a veia jugular externa através do manguito e oclusão do vaso proximal, colocando um grampo vascular sobre a extensão do punho. Remova a ligadura e irrigar o lúmen do vaso com 01:10 salina heparinizada. </li><li> Evert parede do vaso através da braçadeira e fixe-o com uma predefinição circunferencial 8-0 seda ligadura. </li><li> Remova a parte superficial do músculo esternocleidomastóideo. </li><li> Mobilizar a artéria carótida comum direita e transecto embarcação entre ligaduras abaixo da bifurcação carotídea. </li><li> Puxe a artéria carótida através do manguito e oclusão do vaso proximal, colocando um grampo Yasargil sobre a extensão do punho. Remova a ligadura e irrigar o lúmen do vaso com 01:10 salina heparinizada. </li><li> Gentilmente dilate do lúmen arterial, com um dilatador navio, everter parede do vaso através da braçadeira e fixe-o com uma predefinição circunferencial 8-0 seda ligadura. </li><li> Para ganhar espaço suficiente para a implantação do enxerto, remover o lobo direito da glândula submandibular. </li><li> Mantenha o campo operatório úmido até a implantação. </li></ol> 3. Enxerto Implantação <ol><li> Coloque o enxerto na região cervical do receptor de cabeça para baixo com a aorta orientada medialmente e a artéria pulmonar lateralmente. </li><li> Puxe a aorta do enxerto sobre o manguito com a artéria carótida e garantir a anastomose com uma ligadura circunferencial 8-0 seda. </li><li> Inciso do tronco pulmonar no seu aspecto anterior e puxe-o sobre o manguito com a veia jugular externa. Novamente garantir a anastomose com uma ligadura circunferencial 8-0 seda. </li><li> Remover o grampo na veia jugular externa, seguindo por desclampeamento da artéria carótida. Os corações enche imediatamente com sangue. Para aumentar reperfusion cair 37 ° C soro fisiológico morno para o enxerto. Normalmente, o coração se desenvolve em ritmo sinusal dentro de um minuto. Remover o manguito extensão no local venoso. </li><li> Assegurar a posição correta do enxerto, anastomoses deve ser livre de tensão e vasos sem torção. Não vasos cauterizados da pele uma vez que este pode prejudicar a cicatrização de feridas. Sangramento geralmente pára espontaneamente. Fechar a pele com uma sutura contínua única 8-0. </li></ol> 4. Cuidados Pós-Operatórios <ol><li> Injectar 0,25 ml quente sc solução salina para a reposição de líquidos. </li><li> Para antagonizar narcóticos injectar a naloxona (1,2 mg / kg), o flumazenil (0,5 mg / kg) e atipamezol (2,5 mg / kg de peso corporal) SC esquerdo do animal na almofada quente até ter recuperado a consciência suficiente para manter decúbito esternal. Observar o animal durante pelo menos uma hora adicional para registar qualquer comportamento anormal. Não devolva um animal que tenha sido submetido a cirurgia para a companhia de outros animais até que esteja totalmente Recovevermelho. </li><li> Fornecer analgesia preemptiva com buprenorfina (0,05 mg / kg sc três vezes por dia) ou uma analgesia alternativa de acordo com um protocolo institucional local por um período mínimo de 72 horas após o procedimento. </li><li> O cirurgião e um veterinário deve ver os animais regularmente. Em caso de perda de peso&gt; 15%, a apatia, inchaço ou infecção lado cirúrgicos animais são sacrificados usando inalação de isoflurano terminal. </li></ol>

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+obliterative+bronchiolitis+in+a+murine+model+of+orthotopic+lung+transplantation.">Development of obliterative bronchiolitis in a murine model of orthotopic lung transplantation.</a> J Vis Exp : JoVE. , (2012).</li><li>Zou, 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=Mouse+model+of+venous+bypass+graft+arteriosclerosis.">Mouse model of venous bypass graft arteriosclerosis.</a> Am J Pathol. 153, 1301-1310 (1998).</li><li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant.">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Exp Clin Transplant. 8, 245-249 (2010).</li><li>Niimi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+technique+for+heterotopic+cardiac+transplantation+in+mice:+experience+of+3000+operations+by+one+surgeon.">The technique for heterotopic cardiac transplantation in mice: experience of 3000 operations by one surgeon.</a> J Heart Lung Transplant. 20, 1123-1128 (2001).</li><li>Arras, M., Autenried, P., Rettich, A., Spaeni, D., Rulicke, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+intraperitoneal+injection+anesthesia+in+mice:+drugs,+dosages,+adverse+effects,+and+anesthesia+depth.">Optimization of intraperitoneal injection anesthesia in mice: drugs, dosages, adverse effects, and anesthesia depth.</a> Comp Med. 51, 443-456 (2001).</li></ol>

The authors have nothing to disclose.

Nós completamos mais de 200 transplantes cardíacos com uma taxa de sucesso superior a 90% em várias configurações experimentais. Com a técnica descrita aqui, o procedimento completo pode ser feito no prazo de 60 min por um cirurgião experiente. Ao preparar os vasos cervicais antes de colheita de órgãos, o tempo total de isquemia do enxerto pode ser limitada a 20 min. Tempo para o implante (tempo de isquemia quente) é significativamente mais curta utilizando a técnica de braçadeira em comparação com a técn...

O modelo de transplante cardíaco é freqüentemente usada para investigar I / R-lesão, rejeição do enxerto, doença vascular pós-transplante ea eficiência dos agentes imunomoduladores. Mouse modelos combinam muitas vantagens como o imunológica conhecida e origens genéticas, a disponibilidade de muitas estirpes puras e transgénicas experimentais e custos relativamente baixos. Em 1973, uma técnica de transplante cardíaco abdominal em ratinhos foi descrita pela primeira vez por Corry et al 1. Neste modelo enxertos são revascularizada pelo ponta-a-lado anastomose da aorta ascendente a aorta abdominal do receptor e da principal artéria pulmonar à veia cava inferior. Em 1991, a técnica de sutura para o transplante cardíaco cervical em ratos foi descrita por Chen et al 2. Neste modelo a perfusão coronária retrógrada é estabelecido por anastomose da artéria carótida do destinatário à aorta ascendente do grpopa. Os drenos de sangue venoso, através do seio coronário no átrio direito, ventrículo direito e é ejetado através da artéria pulmonar para o destinatário veia jugular externa (Figura 1). Em comparação com a implantação do enxerto abdominal modelo cervical é menos invasiva ea localização superficial do enxerto permite um acesso muito fácil para exames de acompanhamento adicionais (por exemplo palpação, ecocardiografia, microscopia de fluorescência intravital) 3,4. Apesar das melhorias técnicas 5,6, uma utilização mais generalizada deste modelo tem sido limitada devido a dificuldades técnicas para realizar anastomoses suturadas de pequenos vasos com alta incidência de vazamento de anastomose e trombose. Em 1991, Matsuura et ai. introduziu o manguito técnica em que a parede do vaso é revirado sobre um cilindro de síntese, o que facilita grandemente o modelo de transplante de coração 7 do colo do útero. A técnica foi further adotado para vários modelos experimentais de transplante renal, incluindo 8, 9 pancreático, membro 10, hepáticos 11,12, 13,14 e transplante de pulmão vascular 15. Em comparação com a técnica de sutura o tempo necessário para a anastomose e, assim, o tempo de isquemia quente é mais curto e existem complicações vasculares significativamente menor, utilizando a técnica do manguito 16. Além disso, ele permite que os cirurgiões com pouco treinamento em microcirurgia para executar a operação com uma elevada taxa de sucesso. Uma desvantagem da técnica de braçadeira é pré-requisito para a ligação da artéria carótida comum, o que pode alterar a perfusão cerebral. Neste artigo, apresentamos um vídeo técnica manguito melhorado e simplificado para o transplante cardíaco cervical em camundongos.

<table><tbody><tr><td>Forceps JFC-7.2</td><td>S&amp;amp;T</td><td>P-00036</td><td>Curved tip 0.20 mm</td></tr><tr><td>Vessel dilatator D-5a</td><td>S&amp;amp;T</td><td>S-00124</td><td></td></tr><tr><td>Adventita scissor</td><td>S&amp;amp;T</td><td>S-00102</td><td></td></tr><tr><td>Vascular clamp B-1V</td><td>S&amp;amp;T</td><td>S-00396</td><td></td></tr><tr><td>Yasargil Clip</td><td>Peter Lazic</td><td>65,097</td><td>3.5 mm lenght</td></tr><tr><td>Bipolar forceps</td><td>Micromed</td><td>148-100-011</td><td>tip 0.25 mm</td></tr><tr><td>Polyimide tubes</td><td>River Tech Medical</td><td></td><td>19-24 gauge</td></tr><tr><td>8.0 silk ligatures</td><td>Catgut</td><td></td><td></td></tr><tr><td>Custodiol HTK solution</td><td>Essential Pharmaceuticals</td><td></td><td></td></tr></tbody></table>

heterotopic cervical heart transplantation in mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

Nossa equipe pode executar com êxito a técnica descrita após cerca de 20 cirurgias de formação. A parte mais desafiadora é a everter parede do vaso da artéria carótida sobre o punho, porque facilmente vira para trás e múltiplas tentativas podem levar a rasgar do muro. Após o período de treinamento, o tempo cirúrgico médio total foi de 60 ± 8 min. Cerca de 20 min são necessários para a colheita doador, 25 min para a preparação dos vasos cervicais e 15 minutos para anastomoses e fechamento da pele. ...

Watch this Scientific Journal Video about Heterotopic Cervical Heart Transplantation in Mice at JoVE.com

Heterotopic Cervical Heart Transplantation in Mice

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Heterotopic Cervical Transplante Cardíaco em Ratos

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

heterotopic-cervical-heart-transplantation-in-mice

Research

JoVE Journal

Medicine

11.1K visualizações.  German Heart Center Munich, Technische Universität München. The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Vídeo: Heterotopic Cervical Heart Transplantation in Mice

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

Transplante cardíaco heterotópico em ratos

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

Biologia

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

Murino Cervical Modelo de coração Transplante usando uma técnica Cuff Modificado

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

Medicina

A Modified Method for Heterotopic Mouse Heart Transplantion

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents available. A difficulty is presented due to the small size of the animal and the considerable technical challenges of the microsurgery involved in heart transplantation. In particular, a high rate of technical failure early after transplantation may result from recipient death and post-operative complications such as hind limb paralysis or a non-beating heart. Here, the complete technique for heterotopic mouse heart transplantation is demonstrated, involving harvesting the donor heart and its subsequent implantation into a recipient mouse. The donor heart is harvested immediately following in situ perfusion with cold heparinized saline and transection of the ascending aorta and pulmonary artery. The recipient operation involves preparation of the abdominal aorta and inferior vena cava (IVC), followed by end-to-side anastomosis of the donor aorta with the recipient aorta using a single running 10-0 microsuture and a similar anastomosis of the donor pulmonary artery with the recipient IVC. Following the operation the animal is injected with 0.6 ml normal saline subcutaneously and allowed to recover on a 37&#160;&#176;C heating pad. The results from 227 mouse heart transplants are summarized with a success rate at 48 hr of 86.8%. Of the 13.2% failures within 48 hr, 5 (2.2%) experienced hind limb paralysis, 10 (4.4%) had a non-beating heart due to graft ischemic injury and/or thrombosis, while 15 (6.6%) died within 48 hr.

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

Um método modificado para heterotópico Rato coração transplante

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

Técnica de transplante heterotópico Murino Coração

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

A Microsurgical Modelo Novel para heterotópico, em bloco da parede torácica, Thymus, e Transplante Cardíaco em Ratos

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

An Immunological Model for Heterotopic Heart and Cardiac Muscle Cell Transplantation in Rats

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications have been reported since the first description in 1964. After 30 years of performing heterotopic heart transplantation in rats, we have developed a simplified surgical approach, which can be easily taught and performed without further surgical training or background.
After dissection of the ascending aorta and the pulmonary artery and ligation of superior and inferior caval and pulmonary veins, the donor heart is harvested and subsequently perfused with ice-cold saline solution supplemented with heparin. After clamping and incising the recipient abdominal vessels, the donor ascending aorta and pulmonary artery are anastomosed to the recipient abdominal aorta and inferior vena cava, respectively, using continuous running sutures.
Depending on different donor-recipient combinations, this model allows analyses of either acute or chronic rejection of allografts. The immunological significance of this model is further enhanced by a novel approach of in-ear injection of vital cardiac muscle cells and subsequent analysis of draining cervical lymphatic tissue.

We describe a model of heterotopic abdominal heart transplantation in rats, implying modifications of current strategies, which lead to a simplified surgical approach. Additionally, we describe a novel rejection model by in-ear injection of vital cardiac muscle cells, allowing further transplant immunological analyses in rats.

Um modelo imunológico para transplante heterotópico de células cardíacas e cardíacas em ratos

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications ...

Imunologia e Infecção

Optimization of the Cuff Technique for Murine Heart Transplantation

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

Otimização da Técnica de Manguito para Transplante de Coração de Murine

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to ...

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 ...

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

Transplante cardíaco cervical heterotópico de camundongo utilizando manguitos vasculares

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

Reconstrução do circuito sanguíneo em um modelo de transplante de coração de rato abdominal

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...