Questo lavoro è stato sostenuto, in parte, da una sovvenzione del NIH (RO1 HL098228) per CKB.

Nota: Tutte le procedure sugli animali sono state approvate dalla Hospital Institutional Animal Care ed uso Comitato dei Bambini di Nationwide. 1. Polmonare valvola cardiaca raccolta da un donatore Valvola cardiaca mouse <ol><li> Sterilizzare in autoclave tutti gli strumenti chirurgici prima della chirurgia: 1x forbici sottili, 3x micro pinze, pinze vascolari micro 2x, 1x morsetto pinze applicano, porta-aghi micro 1x, forbici primavera 1x, 1x divaricatore. </li><li> A 6-8 settimane di età femmina C57BL / 6 mouse viene utilizzato come donatore valvola cardiaca polmonare. Rimuovere il mouse dalla gabbia e si pesa, poi eutanasia con un cocktail di ketamina / xylazina (ketamina, 200 mg / kg e xylazina, 20 mg / kg, IP) overdose. </li><li> Agganciare la zona del torace e posizionare il mouse in una posizione prona dorsale su un taccuino. Poi fare la toracotomia. Esporre il cuore, fare un piccolo taglio sul atrio destro e profumato il ventricolo sinistro con ghiaccio soluzione fisiologica fredda. </li><li> Senza mezzi termini sezionare l&#39;arteria polmonare (PA) dal crescente unorta. Tagliare la valvola polmonare (PV) insieme a due millimetri bracciale dell&#39;arteria polmonare. Smaltire il resto del cuore. </li><li> Conservare il fotovoltaico in soluzione salina (100 unità / ml) di eparina freddo e. Nota: Il PV può essere mantenuto in soluzione per due ore prima del trapianto verso il cuore del donatore. </li></ol> 2. Preparazione donatore di cuore <ol><li> A 6-8 settimane di età femmina C57BL / 6 mouse viene utilizzato come un donatore di cuore. Rimuovere il mouse dalla gabbia e si pesa, poi eutanasia con un cocktail di ketamina / xylazina (ketamina, 200 mg / kg e xylazina, 20 mg / kg, IP) overdose. Questa è una procedura terminale. </li><li> Agganciare la zona del torace e posizionare il mouse in una posizione prona dorsale su un taccuino. Poi fare la toracotomia. Senza mezzi termini separare il cuore, vena cava inferiore (IVC), vena cava superiore (SVC), dell&#39;aorta ascendente, PA, e la vena polmonare. Profumato l&#39;IVC con il freddo ghiaccio soluzione salina sterile. </li><li> Legare l&#39;IVC, SVC, e vena polmonare con 6-0 sutura di seta poi tagliare superiore allegature. </li><li> Tagliare l&#39;aorta e PA 2 mm bracciale. </li><li> Tagliare il PV e smaltirlo. </li></ol> 3. Valvola cardiaca trapianto su un donatore di cuore <ol><li> Subito dopo la fase 2.5, posizionare la valvola cardiaca dal punto 1.5 nel cuore del donatore e orientare la valvola cardiaca. </li><li> Fissare la PV con un punto sul lato destro della valvola attraverso 10-0 monofilamento sugli aghi conici e inizia a suturare continuamente con 5-6 stiches dall&#39;altro lato del PV. </li><li> Dopo aver terminato il lato anteriore, ruotare cuore orizzontalmente e iniziare a suturare il lato posteriore del PV sul cuore del donatore. </li><li> Conservare il cuore in una soluzione fredda di eparina / soluzione salina sterile. Nota: Il cuore del donatore può essere mantenuto in soluzione per due ore prima dell&#39;impianto per il mouse destinatario. </li></ol> 4. Eterotopico trapianto cardiaco a un destinatario mouse <ol><li> A 6-8 settimane di età femmina C57BL / 6 topo è stato usato come un recipient. Rimuovere il mouse dalla gabbia e si pesa, poi anestetizzati con cocktail di ketamina / xilazina (ketamina, 100 mg / kg e xilazina 10 mg / kg). Ketoprofene (5 mg / kg) è usato come preanesthesia analgesico. </li><li> Dopo aver verificato il livello di sedazione per coda pizzicare, tagliare il addominale e peli del torace. Lubrificare gli occhi con pomata oftalmica sterile, e posizionare il mouse in una posizione prona dorsale su un taccuino. Disinfettare l&#39;addome con Betadine e alcool pastiglie. Poi coprire il mouse con un telino sterile ed esporre solo la zona di incisione. </li><li> Effettuare una laparotomia mediana incisione da sotto la xifoidea alla regione sovrapubica, e inserire un divaricatore auto ritegno. Avvolgere l&#39;intestino in soluzione salina inumidito garza. Senza mezzi termini definire l&#39;aorta sottorenale e la vena cava. </li><li> Inserire due 6-0 punti di sutura in seta prossimale e distale attorno alla aorta e IVC per limitare la circolazione del sangue. </li><li> Posizionare il cuore del donatore sul lato destro dell&#39;aorta addominale e coprire con gau sterileze. Idratare con soluzione fisiologica. </li><li> Effettuare una aortotomy nell&#39;aorta addominale utilizzando un ago G 30 ed estendere l&#39;apertura con le forbici alle dimensioni dell&#39;aorta donatore. </li><li> Eseguire un end-to side anastomosi con sterili suture monofilamento 10-0 sugli aghi conici. Fissare l&#39;aorta donatore con un punto sull&#39;estremità prossimale dell&#39;apertura nell&#39;aorta addominale e inizia a suturare continuamente con 4-5 stiches dall&#39;estremità distale dell&#39;aorta addominale. </li><li> Capovolgere il cuore a sinistra, coprire con garza salina infusa e iniziare a suturare continuamente con 4-5 stiches dall&#39;estremità distale dell&#39;aorta addominale. </li><li> Effettuare una venotomia nella IVC usando un ago G 30 ed estendere l&#39;apertura alla dimensione dell&#39;arteria polmonare donatore. </li><li> Eseguire la fine anastomosi lato utilizzando sterili suture monofilamento 10-0 sugli aghi conici. Fissare il donatore PA con un punto sull&#39;estremità prossimale dell&#39;apertura nella IVC e inizia a suturare continuamente con 4-5 stiches dal&#39;estremità distale della vena cava inferiore. Questa volta, perché l&#39;aorta è nel senso, si assicura la sutura della parete sinistra della PA del donatore è all&#39;interno della IVC. </li><li> Lavare lume IVC con eparina e soluzione salina (100 unità / ml). Chiudere la parete destra del donatore e del ricevente PA IVC suturando loro continuamente all&#39;estremità distale. </li><li> Togliere la legatura distale e controllare l&#39;emorragia applicando una topica agente emostatico sterile riassorbibile. Quando l&#39;emorragia si arresta completamente, rimuovere la sutura prossimale e controllare l&#39;emorragia allo stesso modo. </li><li> Ritorno intestini e chiudere la muscolatura addominale e la pelle in due strati con un 6-0 nero poliammide monofilamento di sutura. </li><li> Iniettare 0,5 ml di soluzione fisiologica per via sottocutanea e posizionare il mouse in una gabbia di recupero su un rilievo di riscaldamento fino a quando il mouse è completamente mobile. Al momento del recupero, ritorno il mouse in una nuova gabbia con biancheria da letto di carta. Dare farmaci antidolorifici (ibuprofene, 30 mg / kg, acqua potabile) per 48 ore. FareNon restituire un animale che ha subito un intervento chirurgico per la compagnia di altri animali fino alla completa guarigione. </li></ol>

<ol>
	<li>Polito, A., 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=Increased+morbidity+and+mortality+in+very+preterm+VLBW+infants+with+congenital+heart+disease.">Increased morbidity and mortality in very preterm VLBW infants with congenital heart disease.</a> Intens Care Med. 39, 1104-1112 (2013).</li><li>Wren, C., Reinhardt, Z., Khawaja, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twenty+year+trends+in+diagnosis+of+life+threatening+neonatal+cardiovascular+malformations.">Twenty year trends in diagnosis of life threatening neonatal cardiovascular malformations.</a> Arch Dis Child Fetal. 93, F33-F35 (2008).</li><li>Vacanti, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+transplantation+Third+annual+Samuel+Jason+Mixter+lecture.">Beyond transplantation Third annual Samuel Jason Mixter lecture.</a> Archives of surgery. 123, 545-549 (1960).</li><li>Tudorache, I., 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=Orthotopic+replacement+of+aortic+heart+valves+with+tissue-engineered+grafts.">Orthotopic replacement of aortic heart valves with tissue-engineered grafts.</a> Tissue engineering Part A. 19, 1686-1694 (2013).</li><li>van Geldorp, M. W., 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=Patient+outcome+after+aortic+valve+replacement+with+a+mechanical+or+biological+prosthesis+weighing+lifetime+anticoagulant+related+event+risk+against+reoperation+risk.">Patient outcome after aortic valve replacement with a mechanical or biological prosthesis weighing lifetime anticoagulant related event risk against reoperation risk.</a> The Journal of thoracic and cardiovascular surgery. 137, 881-886 (2009).</li><li>El Oakley, R., Kleine, P., Bach, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choice+of+prosthetic+heart+valve+in+today's+practice.">Choice of prosthetic heart valve in today's practice.</a> Circulation. 117, 253-256 (2008).</li><li>Quinn, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Bench+to+Bedside+Translation+of+Regenerative+Cardiac+Constructs.">Animal Models for Bench to Bedside Translation of Regenerative Cardiac Constructs.</a> Progress in Pediatric cardiology. 35, (2013).</li><li>Shinoka, 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=Tissue+engineering+heart+valves+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineering heart valves valve leaflet replacement study in a lamb model.</a> The Annals of thoracic surgery. 60, S513-S516 (1995).</li><li>Sodian, R., 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=Tissue+engineering+of+heart+valves+in+vitro+experiences.">Tissue engineering of heart valves in vitro experiences.</a> The Annals of thoracic surgery. 70, 140-144 (2000).</li><li>Shinoka, 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=Tissue+engineered+heart+valves.+Autologous+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineered heart valves. Autologous valve leaflet replacement study in a lamb model.</a> Circulation. 94, II164-II168 (1996).</li><li>Sodian, R., 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=Evaluation+of+biodegradable+three+dimensional+matrices+for+tissue+engineering+of+heart+valves.">Evaluation of biodegradable three dimensional matrices for tissue engineering of heart valves.</a> ASAIO journal (American Society for Artificial Internal Organs. 46, 107-110 (1992).</li><li>Sodian, R., 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=Early+in+vivo+experience+with+tissue-engineered+trileaflet+heart+valves).">Early in vivo experience with tissue-engineered trileaflet heart valves).</a> Circulation. 102, III22-III29 (2000).</li></ol>

Non abbiamo nulla da rivelare.

Il tasso di mortalità di questa procedura è vicino al 20%, che è stato causato principalmente da emorragia nel sito di trapianto PV e anastomosi sull&#39;aorta donatore all&#39;aorta addominale destinatario. Nella maggior parte dei casi, il tasso di mortalità diminuisce significativamente chirurgia 48 hr postale. I topi hanno mostrato di sopravvivenza cuore batte forte e il flusso di sangue attraverso il fotovoltaico impiantato. L&#39;intero processo dura quattro ore per una micro chirurgo esperto. Ci vorranno circa...

Difetti cardiovascolari congenite sono una delle principali cause di mortalità infantile del 1,2 mondo occidentale. Tra questi, la stenosi valvolare polmonare e premolari difetti valvolari aortici sono una forma che si verificano di frequente 3. Intervento di sostituzione della valvola cardiaca è una scelta di routine di chirurgia ricostruttiva; Tuttavia, le complicanze, tra cui la stenosi e calcificazione della valvola cardiaca, e la dipendenza per tutta la vita su anticoagulanti sono una fonte significativa di malattie croniche e morte 4-7. Inoltre, la mancanza di potenziale di crescita richiede interventi di revisione, che aumenta ulteriormente la mortalità di questi pazienti giovani 4,8,9. Nel tentativo di sviluppare una valvola cardiaca sostituzione funzionale con potenziale di crescita, le cellule autologhe seminati Shinoka et al. Su una valvola cardiaca sintetico biodegradabile 8. La valvola sintetico trasformato in una valvola cardiaca nativa come la struttura con la crescita potenziometroal. Preliminari studi su animali di grandi dimensioni hanno dimostrato la possibilità di utilizzare questa metodologia per creare una valvola cardiaca funzionale 10. Tuttavia, gli studi di impianto lungo termine hanno dimostrato scarsa durata a causa del progressivo ispessimento della neotissue valvola con conseguente restringimento della valvola cardiaca. Lavorare da Sodian et al. Utilizzata la metodologia Shinoka, ma alla fine sostituito matrice PGA con un elastomero biodegradabile, che ha le proprietà biomeccaniche della valvola di ingegneria tessutale costruire un profilo più fisiologico 9,11,12. Nello studio in vivo, nonostante il successo del l&#39;impianto, un rivestimento delle cellule endoteliali confluenti non era formata che potrebbe limitare il successo a lungo termine di questo ponteggio 12. Al fine di progettare razionalmente una migliore valvola cardiaca sintetico di seconda generazione, un modello murino di trapianto di valvole cardiache è stato creato per studiare i meccanismi cellulari e molecolari underlyinformazione g neotissue, ispessimento della valvola, e lo sviluppo di stenosi. Modelli murini offrono una vasta gamma di reagenti molecolari, tra cui transgenici, che non sono facilmente disponibili in altre specie 7. In questo cuore modello di trapianto di valvole, un ex vivo singenico sostituzione della valvola cardiaca polmonare è stata eseguita prima; e poi il cuore con la valvola cardiaca impiantato è stato impiantato heterotopically in un ospite singenici utilizzando una tecnica microchirurgica. Questo modello permette la sostituzione della valvola cardiaca senza la necessità di bypass cardiopolmonare. In questo lavoro, una spiegazione dettagliata di un raccolto valvola cardiaca, preparazioni cuore del donatore, il trapianto di valvole cardiache, e il trapianto di cuore eterotopico è descritto. I risultati hanno mostrato un battito cardiaco continuo dal cuore del donatore, che era indipendente dal battito cardiaco destinatario. Il flusso sanguigno attraverso la valvola polmonare impiantato è stata misurata usando un sistema a ultrasuoni ad alta frequenza con onda pulsata Fareppler.

<table><tbody><tr><td>DPBS</td><td>Gibco</td><td>14190-144</td><td></td></tr><tr><td>Microscope</td><td>Leica</td><td>M80</td><td></td></tr><tr><td>C57BL/6J (H-2b), Female</td><td>Jackson Laboratories</td><td>664</td><td>8-12 weeks</td></tr><tr><td>Ketamine Hydrochloride Injection</td><td>Hospira Inc.</td><td>NDC 0409-2053</td><td></td></tr><tr><td>Xylazine Sterile Solution</td><td>Akorn Inc.</td><td>NADA# 139-236</td><td></td></tr><tr><td>Ketoprofen</td><td>Fort Dodge Animal Health</td><td>NDC 0856-4396-01</td><td></td></tr><tr><td>Ibuprofen</td><td>PrecisionDose</td><td>NDC 68094-494-59</td><td></td></tr><tr><td>Heparin Sodium</td><td>Sagent Pharmaceticals</td><td>NDC 25021-400</td><td></td></tr><tr><td>Saline solution (Sterile 0.9% sodium chloride)</td><td>Hospira Inc.</td><td>NDC 0409-0138-22</td><td></td></tr><tr><td>0.9% Sodium chloride Injection</td><td>Hospira Inc.</td><td>NDC 0409-4888-10</td><td></td></tr><tr><td>Petrolatum Ophthalmic Ointment</td><td>Dechra Veterinary Products</td><td>NDC 17033-211-38</td><td></td></tr><tr><td>Iodine Prep Pads</td><td>Triad Disposables, Inc.</td><td>NDC 50730-3201-1</td><td></td></tr><tr><td>Alcohol Prep Pads</td><td>McKesson Corp.</td><td>NDC 68599-5805-1</td><td></td></tr><tr><td>Cotton tipped applicators</td><td>Fisher Sientific</td><td>23-400-118</td><td></td></tr><tr><td>Fine Scissor</td><td>FST</td><td>14028-10</td><td></td></tr><tr><td>Micro-Adson Forcep</td><td>FST</td><td>11018-12</td><td></td></tr><tr><td>Clamp Applying Forcep</td><td>FST</td><td>00072-14</td><td></td></tr><tr><td>S&amp;amp;T Vascular Clamp</td><td>FST</td><td>00396-01</td><td></td></tr><tr><td>Spring Scissors</td><td>FST</td><td>15008-08</td><td></td></tr><tr><td>Colibri Retractors</td><td>FST</td><td>17000-04</td><td></td></tr><tr><td>Dumont #5 Forcep</td><td>FST</td><td>11251-20</td><td></td></tr><tr><td>Dumont #7 - Fine Forceps</td><td>FST</td><td>11274-20</td><td></td></tr><tr><td>Dumont #5/45 Forceps</td><td>FST</td><td>11251-35</td><td></td></tr><tr><td>Tish Needle Holder/Forceps</td><td>Micrins</td><td>MI1540</td><td></td></tr><tr><td>Black Polyamide Monofilament Suture, 10-0</td><td>AROSurgical Instruments Corporation</td><td>TI638402</td><td>For sutureing the graft</td></tr><tr><td>Black Polyamide Monofilament Suture, 6-0</td><td>AROSurgical Instruments</td><td>SN-1956</td><td>For musculature and skin closure</td></tr><tr><td>Non Woven Sponges</td><td>McKesson Corp.</td><td>94442000</td><td></td></tr><tr><td>Absorbable hemostat</td><td>Ethicon</td><td>1961</td><td></td></tr><tr><td>1 ml Syringe</td><td>BD</td><td>309659</td><td></td></tr><tr><td>3 ml Syringe</td><td>BD</td><td>309657</td><td></td></tr><tr><td>10 ml Syringe</td><td>BD</td><td>309604</td><td></td></tr><tr><td>18 G 1 1/2 in, Needle</td><td>BD</td><td>305190</td><td></td></tr><tr><td>25 G 1 in., Needle</td><td>BD</td><td>305125</td><td></td></tr><tr><td>30 G 1 in., Needle</td><td>BD</td><td>305106</td><td></td></tr><tr><td>Warm Water Recircultor</td><td>Gaymar</td><td>TP-700</td><td></td></tr><tr><td>Warming Pad</td><td>Gaymar</td><td>TP-22G</td><td></td></tr><tr><td>Trimmer</td><td>Wahl</td><td>9854-500</td><td></td></tr><tr><td>VEVO2100 High Frequency Ultrasound</td><td>VisualSonics</td><td>&lt;a href=&quot;http://www.visualsonics.com/vevo2100&quot;&gt;http://www.visualsonics.com/vevo2100&lt;/a&gt;</td><td>The catalog number and pricing can be acquired from the sales representatives.</td></tr><tr><td>Ultrasound transmission gel</td><td>Parker Laboratories,&lt;br/&gt; INC.</td><td>01-02</td><td></td></tr><tr><td>Table Top Laboratory Animal Anesthesia System</td><td>VetEquip, INC.</td><td>901806</td><td></td></tr><tr><td>Isoflurane</td><td>Baxter</td><td>1001936060</td><td></td></tr></tbody></table>

transplantation of pulmonary valve using a mouse model of heterotopic heart transplantation

Valvole cardiache di ingegneria tessutale, valvole soprattutto decellularized, stanno iniziando a guadagnare slancio in uso clinico di chirurgia ricostruttiva con risultati alterni. Tuttavia, i meccanismi cellulari e molecolari dello sviluppo neotissue, ispessimento della valvola, e lo sviluppo stenosi non sono studiati estesamente. Per rispondere a queste domande, abbiamo sviluppato una valvola cardiaca modello murino di trapianto eterotopico. Una valvola cardiaca è stato raccolto da un topo donatore valvola e trapiantato in un topo donatore di cuore. Il cuore con una nuova valvola è stato trapiantato heterotopically ad un mouse destinatario. Il cuore trapiantato ha mostrato il proprio battito cardiaco, indipendente battito cardiaco del destinatario. Il flusso di sangue è stato quantificato usando un sistema a ultrasuoni ad alta frequenza con un Doppler pulsato. Il flusso attraverso la valvola polmonare impiantata mostrato flusso in avanti con rigurgito minimo e il picco di flusso era vicino a 100 mm / sec. Questo modello murino di trapianto di valvola cardiaca è highly versatile, in modo che possa essere modificato e adattato per fornire diversi ambienti emodinamici e / o può essere usato con vari topi transgenici per studiare lo sviluppo neotissue in una valvola cardiaca ingegneria tessutale.

La Figura 1 illustra gli schemi della valvola modello di trapianto di cuore con trapianto di cuore eterotopico. La valvola cardiaca è stato raccolto da un donatore e impiantato su un cuore da un secondo mouse donatore. Poi il cuore con la nuova valvola cardiaca è stato impiantato all&#39;addome di un mouse destinatario. Figura 2 mostra una illustrazione del cuore impiantato nello spazio addominale (A), subito dopo trapianto cardiaco (B), e 5 minuti do...

Watch this Scientific Journal Video about Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation at JoVE.com

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

Al fine di comprendere i meccanismi cellulari e molecolari alla base neotissue formazione e lo sviluppo di stenosi in ingegneria tessutale valvole cardiache, è stato sviluppato un modello murino di trapianto di valvole cardiache eterotopico. Una valvola cardiaca polmonare è stato trapiantato al destinatario utilizzando la tecnica di trapianto di cuore eterotopico.

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 ...

transplantation-pulmonary-valve-using-mouse-model-heterotopic-heart

Research

JoVE Journal

Medicine

18.8K Views.  Nationwide Children's Hospital.  Al fine di comprendere i meccanismi cellulari e molecolari alla base neotissue formazione e lo sviluppo di stenosi in ingegneria tessutale valvole cardiache, è stato sviluppato un modello murino di trapianto di valvole cardiache eterotopico. Una valvola cardiaca polmonare è stato trapiantato al destinatario utilizzando la tecnica di trapianto di cuore eterotopico. 

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

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.

Il trapianto cardiaco eterotopico nel 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 ...

Ricerca

Biologia

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 ...

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 ...

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 ...

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 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 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 ...

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.

Trapianto cardiaco cervicale eterotopico di topo utilizzando bracciali vascolari

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

Immunologia e infezioni

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.

Ricostruzione del circuito sanguigno in un modello di trapianto di cuore di topo addominale

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

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 Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

Una tecnica di bracciale modificata per il modello di trapianto di cuore eterotopico cervicale del topo

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...