This work was funded by the American Association of Plastic Surgeons 2014 Academic Scholar Award.

Tutte le procedure operative sono state completate in conformità con la Johns Hopkins University e il Dipartimento dell&#39;Agricoltura degli Stati Uniti e le esigenze di servizio di sanità pubblica. Questo protocollo segue il Hopkins University cura e l&#39;uso degli animali Comitato Johns, comitato istituzionale di revisione approvato le linee guida (numero di protocollo M013M490). Dati di sopravvivenza finale è stato registrato per le procedure chirurgiche descritte di seguito. Entrambi gli animali donatori e beneficiari ricevono l&#39;anestesia di prelazione con buprenorfina a 0.1 mg / kg sc un&#39;ora prima di un intervento chirurgico e la buprenorfina destinatario animale è ri-somministrato alla stessa dose dopo il trapianto e ri-dosato come necessario nelle prime 48 ore dopo l&#39;intervento chirurgico. 1. donatori Allograft recupero Nota: Inizia la parte del donatore del trapianto 40 minuti prima del trapianto destinatario di minimizzare il tempo di anestesia destinatario e per facilitare un tempo di fine simultaneo o leggermente orecchioora di fine lier contro la preparazione destinatario. <ol><li> Utilizzare strumenti di microchirurgia sterili standard e guanti sterili per la procedura. Il nostro laboratorio utilizza la sterilizzazione in autoclave di strumenti di microchirurgia. </li><li> Anestetizzare il mouse donatore (maschio) con isoflurano vaporizzatore induzione al 4%. Uso Clippers meccanici atraumatica rimuovere i capelli dalla cervicale, toracica, addominale e della regione. Posto l&#39;animale in posizione supina e mantenere isoflurane sul 1-2% attraverso un cono naso. Garantire un&#39;adeguata anestesia durante tutta la procedura, valutando periodicamente il riflesso di punta di ritiro pizzico. </li><li> Prima l&#39;incisione cutanea, ampiamente preparare il dispositivo applicando iodopovidone antisettico seguito da alcol isopropilico utilizzando un tampone di cotone sterile. </li><li> Inizia con una incisione cutanea superficiale trasversale con le forbici in tutta la pelle cervicale e addominale. Collegare i due incisioni bilateralmente lungo le linee ascellare media. </li><li> Utilizzando pinze microchirurgichesezionare la regione cervicale bilaterale per identificare, legare e dividere le vene giugulari esterne con 6-0 sutura di seta e forbici. Quindi, utilizzando elettrocauterizzazione dividere i muscoli sternocleidomastoideo per esporre le vene giugulari interne e le arterie carotidi comuni, a livello bilaterale. Passare una sutura 6-0 di seta sotto il lato sinistro e il lato destro carotide comune e le vene giugulari interne in modo sfuso. NOTA: Saranno legate e divise in seguito al passaggio 1.9. </li><li> Acutamente dividere i muscoli cinghia e associati tessuto areolare sciolto, che si trova anteriori alla trachea, con le forbici per liberare i restanti allegati della regione cervicale. </li><li> Utilizzando elettrocauterizzazione bipolare e la dissezione tagliente, dividere i pettorale muscoli principali e le clavicole per esporre i vasi succlavi e legare (6-0 sutura di seta) e dividere prossimale. </li><li> Poi, delicatamente, afferrare ed estrarre il pene dell&#39;animale. Lungo il dorso del pene visualizzare la vena dorsale del pene, e disinfettare laregione con alcol isopropilico. Utilizzando un ago 30 G, iniettare 30.000 unità di eparina per via endovenosa attraverso la vena dorsale e permettono al pene di rinculo indietro alla sua posizione originale. Può verificarsi perdite parziale della soluzione di eparina nel tessuto circostante. </li><li> Utilizzando i legami di massa precedentemente posizionati intorno carotide comune e la vena giugulare interna, legare e dividere le strutture, bilateralmente. </li><li> Successivo utilizzo forbici per rendere una incisione intraddominale trasversale. Sviscerare l&#39;intestino per esporre la infraepatica vena cava inferiore e iniettare 2 ml di soluzione di cardioplegia fredda Euro-Collins nel infraepatica vena cava inferiore. Garantire la corretta iniezione visualizzando scolorimento del fegato e la cessazione del battito cardiaco prima di passare al passo successivo. NOTA: la soluzione Euro-Collins è preparato nel nostro laboratorio, vedi tabella di reagenti e strumenti specifici. </li><li> Utilizzando le forbici accedono cavità intratoracica tramite un DIAFRAMMA bilateraleincisione ragmatic dall&#39;addome esposta. Estendere l&#39;incisione cefalica attraverso i muscoli intercostali e le costole. Riflettere parete toracica esponendo il cuore, il timo, e grandi vasi assicurando allo stesso tempo la conservazione dei vasi toracici interni lungo la parete toracica. </li><li> Iniettare la sovraepatica vena cava inferiore con 4 ml di soluzione di cardioplegia fredda Euro-Collins. </li><li> Identificare la radice dell&#39;aorta e tracciare distalmente alla aorta discendente. Acutamente tagliare l&#39;aorta discendente (conservando lunghezza massima). </li><li> Identificare il tronco polmonare e dividere appena prossimale al suo punto di filiale (conservando lunghezza massima). Quindi, utilizzando 2 ml di soluzione di cardioplegia fredda Euro-Collins, lavare il tronco polmonare e il cuore mettendo un morbido punta del catetere di plastica nel lume del tronco polmonare. </li><li> Utilizzando una sutura di seta 6-0, legare e sezionare la vena cava inferiore, confluenza delle vene polmonari, e rami accessorie del cava bilaterale cava superiore. Poi elevaree sezionare la cefalica cuore dagli attaccamenti lungo bronchi fusto principale e la trachea con attenzione a non inserire le vie respiratorie. Utilizzando elettrocauterio bipolare acuto e sezionare la parete toracica, il timo, e il cuore completamente liberandolo dal mouse donatore. </li><li> Infine, tagliare la parete toracica allogenico ex vivo a una dimensione inferiore, con le forbici, che attraversa lo sterno e costae laterali, con attenzione a non interrompere i vasi toracici interni (Figura 1A). Per ridurre al minimo emorragia dopo rivascolarizzazione, utilizzare electrocauterization bipolare lungo i confini dello sterno osteomusculocutaneous. </li><li> Posizionare il allogenico in 10 ml di freddo (4 o Celsius) soluzione Euro-Collins se il destinatario non è preparato per incasso. Tuttavia, se il destinatario è pronto per inserto, trasferire l&#39;alloinnesto direttamente al campo operatorio destinatario. </li></ol> 2. Destinatario Preparazione Nota: Per ridurre al minimo il tempo di anestesia destinatario,iniziare la preparazione destinatario in una stazione operativa separata circa 40 minuti prima del completamento del raccolto allograft donatore. <ol><li> Utilizzare un insieme separato di strumenti di microchirurgia sterili standard e guanti sterili per la procedura. </li><li> Anestetizzare il mouse destinatario (maschio o femmina) con isoflurano vaporizzatore induzione al 4%. Utilizzando Clippers meccanici atraumatica rimuovere i capelli dalla regione cervicale e toracica destra. </li><li> Posizionare il mouse in posizione supina e l&#39;angolo l&#39;arto superiore destro leggermente inferiormente formando un angolo di 110 gradi tra la testa e arto superiore destro. Mantenere l&#39;anestesia sul 1-2% isoflurano attraverso un cono naso. </li><li> Posizionare petrolio pomata oftalmica sugli occhi del mouse utilizzando un cotton-fioc. Prima l&#39;incisione cutanea, ampiamente preparare il sito operatorio utilizzando iodopovidone antisettico seguito da alcol isopropilico. </li><li> Utilizzando le forbici, fare una incisione cutanea dalla linea mediana lungo la destra bor inferioreder della mandibola e prolungare l&#39;incisione infero lateralmente alla regione toracica destra. Utilizzando smussa con pinze microvascolari, mobilitare la vena giugulare esterna circonferenziale libero nave da tessuti molli e avventizia. Dividere tutti i rami con elettrocauterizzazione, e rimuovere il lobo destro della ghiandola sottomandibolare utilizzando dissezione tagliente e elettrocauterizzazione di spazio libero per il trapianto allogenico. </li><li> Assicurare una lunghezza sufficiente della vena giugulare esterna per rovesciare su un bracciale, e legare la vena giugulare esterna usando una sutura 6-0 seta. Inserire la vena attraverso il lume di un bracciale pretagliati poliimmide e utilizzare un morsetto microvascolare bulldog per risolvere il complesso nave-polsino in posto. Poi con le forbici, prossimale dividere la vena giugulare esterna, Evert sopra il polsino, e fissare con una sutura in nylon 10-0. (Figura 1B) </li><li> Dividere il muscolo sternocleidomastoideo destro con elettrocauterizzazione bipolare per esporre l&#39;arteria carotide comune. Mob circonferenzialeilize l&#39;arteria cefalica al distale punto più all&#39;interno della regione cervicale. Questa operazione viene eseguita utilizzando scollamento della nave con una pinza per rimuovere tessuti molli e avventizia circostante. </li><li> Utilizzando sutura di seta 6-0, legare e sezionare l&#39;arteria carotide comune. Passare l&#39;arteria attraverso il lume di un bracciale pretagliati poliimmide e fissarlo con un bulldog microvascolare morsetto più vicino possibile al toracico possibile. Dividete il vaso distale, dilatare delicatamente il recipiente con un dilatatore microchirurgico, Evert sopra il polsino, e fissare con una sutura in nylon 10-0. (Figura 1B) NOTA: Il dilatatore microchirurgica specifico è descritto nella tabella di reagenti e strumenti specifici. </li></ol> 3. Allograft dell&#39;inserzione <ol><li> Mantenere la strumentazione sterili standard guanti sterili per posizionare il allotrapianto all&#39;interno della regione cervicale destinatario in un testa in giù e posizione obliqua. </li><li> Quindi, posizionare il aor donatore discendentelumen tic sul costrutto bracciale arteriosa del destinatario e fissarlo con una sutura di nylon 10-0 (Figura 1C e 1D). </li><li> Moda la stessa anastomosi come al punto 3.2 tra l&#39;arteria polmonare donatore e il estroflesso esterna giugulare vena-polsino costrutto del mouse destinatario (Figura 1C e 1D). </li><li> Innanzitutto rimuovere morsetto microvascolare venoso (morsetto vena giugulare esterna) e poi allentare il blocco arterioso (arteria carotide comune morsetto). Durante la riperfusione arteriosa, controllare la totalità del allogenico di affrontare qualsiasi emorragia. Se emorragia viene visualizzato, riapplicare la pinza arteriosa per ridurre al minimo la perdita di sangue e ridurre la fonte di sanguinamento con elettrocauterizzazione bipolare. </li><li> Controllare l&#39;innesto e di garantire l&#39;emostasi. Rilasciare e rimuovere completamente il morsetto microvascolare arteriosa. Osservare il cuore a mostrare segni di riperfusione, che saranno istantaneamente apparente con l&#39;espansione del volume rapida delle camere cardiache, e attendere bmangiare per iniziare entro 0,5-1 min. Utilizzare salina calda (35 ° C) per inumidire il cuore. </li><li> Telo parete toracica in posizione anatomica in modo da non indurre alcuna attorcigliamento o tensioni sui anastomosi. Chiudere la pelle della ferita chirurgica sta usando 6-0 punti di sutura in nylon continui (Figura 1E). </li></ol> 4. Cura postoperatoria <ol><li> Somministrare un 0,3 ml di soluzione salina normale bolo fluido intraperitoneale immediatamente dopo l&#39;intervento per la sostituzione del fluido. </li><li> Poi via sottocutanea iniettare buprenorfina (0,1 mg / kg) e enrofloxacina (5mg / kg) per la profilassi del dolore e infezione, rispettivamente. </li><li> Posto l&#39;animale sotto una lampada di calore fino risveglio dall&#39;anestesia e tornare al decubito sternale. Durante il recupero, ispezionare il collo per visualizzare il battito cardiaco fibrillazione del allotrapianto garantire un&#39;adeguata perfusione allogenico. </li><li> Una volta sveglio e in posizione supina, restituire il mouse in una gabbia separata (senza la compagnia di altri topi) Dove riceva cibo e acqua ad libitum. A causa di qualsiasi movimento restrittiva minore temporanea del arto superiore destro, lasciare una fonte di cibo gelatina sul pavimento della gabbia. </li><li> Osservare il mouse destinatario per 1 ora dopo l&#39;intervento e poi tornare alla struttura gabbia dove possa ricevere cibo e acqua ad libitum e viene ispezionato tre volte al giorno per le prime 24 ore di attività e di apporto nutrizionale. Monitorare i topi per i segni di dolore e di angoscia e ri-dosaggio con buprenorfina (0.1mg / kg) per via sottocutanea due volte al giorno come necessario per la prima 72 ore. Esaminare gli animali ogni giorno da allora in poi e pesare ogni settimana. </li><li> Consultare un membro del personale veterinario, se eventuali topi mostrano segni di dolore, angoscia, o diminuito l&#39;assunzione di cibo. Considerare presto l&#39;eutanasia (nel nostro protocollo la tecnica eutanasia impiega CO 2 overdose per 7 minuti, seguita da dislocazione cervicale). </li><li> Cessazione del battito cardiaco allogenico è definito come un endpoint specifico spingendo il mouse per essere sacrificed. </li></ol>

The authors do not have any conflicts of interest or financial disclosures to declare.

Ci sono una moltitudine di fenomeni che fattore nella ricerca immunologica di allotrapianto, che includono ma non sono limitati a meccanismi di rigetto acuto e cronico, la presentazione diretta e indiretta dell&#39;antigene, destinatario di sensibilizzazione o l&#39;induzione di chimerismo misto. 19 modelli animali sono diventati il gold standard per lo studio di Immunologia dei Trapianti, e modelli di topo sono comunemente implementate a causa del loro basso costo, la disponibilità di transgenici e gene top...

Cardiac transplantation is the treatment of choice for end-stage heart failure. Both technical advancements and pharmacological innovations have propelled the field to early graft acceptance rates above 90%.12,13 Despite this, 60-80% 5-year graft survival is at a standstill and chronic rejection, characterized by transplant vasculopathy, remains inevitable.14-16 Furthermore, patients are subjected to multiple surgical procedures and lifelong immunosuppression, which are associated with chest wall deformities and medical sequelae and toxicities, respectively. The need for innovative approaches to extend allograft survival, minimize the immunosuppressive requirements, and offer reconstructive options for anatomical deformities is pressing.
Vascularized composite allotransplantation offers a unique strategy for improving heart transplant outcomes both from an immunological aspect as well as a reconstructive perspective.17 Vascularized composite allografts are also unique in a way that they have an inherent source of donor-derived hematopoietic stem cells which has shown a favorable ability to reduce immunosuppression and induce and sustain mixed chimerism.1-8 Additionally, co-transplantation of the thymus has shown to prolong survival of both, solid organ transplants and vascularized composite allografts.2,9-11 Combining these strategies with heart transplantation offers a novel solution to the aforementioned challenges facing heart transplantation.18
Murine models serve as excellent platforms for mechanistic in vivo investigation because of the availability of antibodies and well-defined inbred and knockout strains.19-21 Although heart transplantation in mice is commonly studied using a heterotopic intraabdominal microsurgical suture transplant model22-25, a heterotopic, cervical, non-suture cuff technique model has shown to be extremely replicable, reliable, and carries fewer rates of thrombosis.19,26,27 The goal of this study is to develop a heterotopic en bloc osteomyocutaneous chest wall, thymus, and heart transplant technique in mice to study the immunological mechanisms of combined solid organ and vascularized composite allotransplantation using a cervical non-suture cuff technique. This cluster allograft is perfused through the anastomosis of the donor descending aorta to the right common carotid artery and the donor pulmonary artery to the right external jugular vein. Preservation of the internal thoracic vessels and associated thymus branches is paramount to perfusing the chest wall (sternum, ribs, muscles, and skin) and thymus.

<table border="0">
	<tbody>
		<tr>
			<td>Euro-Collins Solution</td>
			<td></td>
			<td></td>
			<td>The solution is not commercially purchased but rather prepared in the laboratory. To make a 500ml solution add the ingredient listed below to a 330ml of double distilled water. Mix well, and then fill in the rest of the 170ml of double distilled water into the solution to a final volume of 500ml. 
			Ingredients: 1.02g KH2PO4, 3.66g K2HPO4, 0.56g KCl, 0.42g NaHCO3, and 17.52g of glucose.</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Ethilon</td>
			<td>MWI 72667</td>
			<td>6-0 Ethilon</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Microsurgery Instruments, Inc.</td>
			<td>10-0 Nylon</td>
			<td>10-0 Nylon</td>
		</tr>
		<tr>
			<td>Polyimide Cuff Vein (21G)</td>
			<td>Vention Medical</td>
			<td>141-0043</td>
			<td>http://www.ventionmedical.com/products-and-services/polyimide-tubing/</td>
		</tr>
		<tr>
			<td>Polyimide Cuff Artery (24G)</td>
			<td>Vention Medical</td>
			<td>141-0027</td>
			<td>http://www.ventionmedical.com/products-and-services/polyimide-tubing/</td>
		</tr>
		<tr>
			<td>Soft plastic tip catheter</td>
			<td>Terumo</td>
			<td>SR*OX2419CA</td>
			<td>24G x 3/4&#34;&#160;</td>
		</tr>
		<tr>
			<td>Microsurgical dilator</td>
			<td>S&#38;T</td>
			<td>D-5a.1</td>
			<td>Dilator, 11cm, FH, 0.1mm AT10d</td>
		</tr>
	</tbody>
</table>

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.

Singenico C57BL / 6 trapianti raggiunto sopravvivenza a lungo termine. Il disegno del eterologo (Figura 1) si sono dimostrati efficaci da una prospettiva di sopravvivenza degli animali e la capacità di valutare la sopravvivenza allograft corso. Questo è stato dimostrato attraverso la pelle sovrastante rimanente praticabile, crescita attiva dei capelli allogenico in corso, e battiti cardiaci sono stati in grado di valutare con la visualizzazione e il dito palpazione. Dati di sopravvivenza è rappresent...

Watch this Scientific Journal Video about A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice at JoVE.com

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

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 Novel microchirurgica modello per eterotopica, in blocco parete toracica, Timo, e trapianto cardiaco nei topi

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

a-novel-microsurgical-model-for-heterotopic-en-bloc-chest-wall-thymus

Research

JoVE Journal

Medicine

9.2K Views.  Johns Hopkins University School of Medicine. 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.

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

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.

Un metodo modificato per eterotopica mouse Cuore Trapianto

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 ...

Ricerca

Medicina

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.

Murini eterotopica cuore Tecnica Trapianto

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 ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

Il trapianto di valvola polmonare Utilizzando un modello murino di trapianto cardiaco eterotopico

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

Rat eterotopico addominale Cuore / Trapianti Single-polmone in una configurazione del volume-caricato

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

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.

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.

Eterotopico cervicale trapianto cardiaco nei topi

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

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

Eterotopico renale Autotrapianto in un modello di suino: un protocollo Step-by-Step

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Cardiopulmonary Bypass in a Mouse Model: A Novel Approach

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization and for minimizing organ damage resulting from prolonged extracorporal circulation. The goal of this paper was to demonstrate a fully functional and clinically relevant model of cardiopulmonary bypass in a mouse. We report on the device design, perfusion circuit optimization, and microsurgical techniques. This model is an acute model, which is not compatible with survival due to the need for multiple blood drawings. Because of the range of tools available for mice (e.g., markers, knockouts, etc.), this model will facilitate investigation into the molecular mechanisms of organ damage and the effect of cardiopulmonary bypass in relation to other comorbidities.

This paper describes how to perform cardiopulmonary bypass in mice. This novel model will facilitate the investigation of the molecular mechanisms involved in organ damage.

Esclusione cardiopolmonare in un modello murino: un nuovo approccio

As prolonged cardiopulmonary bypass becomes more essential during cardiac interventions, an increasing clinical demand arises for procedure optimization ...

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 ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

Un modello semplificato per il trapianto di valvole cardiache eterotopiche nei roditori

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of ...

A Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts and complications such as primary graft dysfunction and graft rejection. The recent clinical use of an ex vivo perfusion device in cardiac transplantation introduces a unique opportunity for treating cardiac allografts with therapeutic interventions to improve function and avoid deleterious recipient responses. Establishing a translational, large-animal model for therapeutic delivery to the entire allograft is essential for testing novel therapeutic approaches in cardiac transplantation. The porcine, heterotopic heart transplantation model in the intraabdominal position serves as an excellent model for assessing the effects of novel interventions and the immunopathology of graft rejection. This model additionally offers long-term survival for the pig, given that the graft is not required to maintain the recipient's circulation. The aim of this protocol is to provide a reproducible and robust approach for achieving ex vivo delivery of a therapeutic to the entire cardiac allograft prior to transplantation and provide technical details to perform a survival heterotopic transplant of the ex vivo perfused heart.

We present a protocol for utilizing a normothermic ex vivo sanguinous perfusion system for the delivery of therapeutics to an entire cardiac allograft in a porcine heterotopic heart transplant model.

Un protocollo di trapianto di cuore eterotopico suino per la somministrazione di terapie a un allotrapianto cardiaco

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...