Mark J. Kearns (Doktorlar ve Kanada Royal College of Surgeons aracılığıyla verilen) UBC Klinisyen Araştırmacı Programı ve UBC 4YF Doktora Bursu destek alır. John H. Boyd Sağlığı Araştırma Scholar için Ulusal Sanatoryum Derneği ve Michael Smith Vakfı. CIHR aracılığıyla Fon. Yazarlar kurulum ve perfüzyon ekipmanın bakım ile yardım için Dr. M. Allard ve Richard Wamboldt teşekkür etmek istiyorum.

Tüm hayvanların barındırıldığı ve Ulusal ve bakım ve laboratuar hayvanlarının kullanımı için kurumsal kurallar ile uyumlu bakım için. Bu protokol Etik onay British Columbia Hayvan Bakım Komitesi Üniversitesi tarafından verildi. 300 arasındaki ağırlığındaki erkek Sprague-Dawley sıçanları - 450 g bu protokol için kullanılmıştır. 1. Donör Operasyonu <ol><li> 3-yollu uzun intravenöz (IV), kateter tüp bağlanan bir silindirik bir şişeye kardiyopleji (RT), yaklaşık olarak 100 ml yazıldı. Yerçekimi tarafından kardiyopleji teslim edilmesini sağlayarak, yaklaşık 80cm operatif yüzeye yukarıda şişeyi yükseltmesine bir stand kullanın. </li><li> Yeterince yapılarını görselleştirmek için, cerrahi loupes bir çift veya bir diseksiyon mikroskobu birini kullanın. NOT: Şu anda 3.4 ile binoküler mikroskop çalışma kullanın - 21.3X büyütme. </li><li> Bir anestezik odasında donör koyun ve 4 ile anestezi neden -% 5 izofluran. </li&gt; <li> Bir işletim platformuna sıçan aktarın ve 1 ile burun konisi ile anestezi korumak -% 2 izofluran. Kuruluğunu önlemek için hayvanın gözlerine veteriner merhem sürün. 25 G iğne ile midazolam (2 mg / kg) intraperitoneal olarak yönetmek. </li><li> Cerrahi makası kullanarak, alt çeneye xiphisternum gelen donör tıraş. (Sonradan IV erişimi için), sol kasık küçük bir bölgesini tıraş. Operatif yüzeylere bir Tüyleri ajanı uygulayın, yaklaşık 5 dakika bekleyin ve gazlı bez parçası ile saç kaldırmak. </li><li> Bir povidon-iyot veya chlorexidine tabanlı bir çözüm olan cerrahi siteleri (biz sadece chlorhexidine kullanın) hazırla. % 0.5 lidokain subkutan - 0.1 ile insizyonel siteleri sızmak. </li><li> Anestezi derinliği ve vital bulguların izlenmesi için ücretsiz sağ hindlimb bırakarak, yapışkan bant ile çalışma platformu ön ayakları ve sol hindlimb sabitleyin. </li><li> Pedal tutam uygun anestezi derinliği olduktan sonra kullanarak, sol kasık kırışık bir kesi yapmak22-bıçak neşter. Künt diseksiyon ile, sol ana femoral ven ortaya çıkarmak ve şöyle IV erişim elde: <ol><li> Yavaşça femoral ven örten doku geri ve 24 G IV damar cannulate. Tuzlu dolu IV boru kısa bir uzunluğa IV bağlayın ve yapışkan bant ile yerine tüp sabitleyin. NOT: sıçanlarda femoral venöz kanülasyon için prosedür Jespersen ve arkadaşları 14 tarafından başka açıklanmıştır. </li><li> IV doğru konumlandırma sağlamak için kan aspire hafifçe IV boru bir tuzlu dolu 10 ml şırınga bağlayın ve. </li><li> IV tüp aracılığıyla fraksiyone olmayan heparine 500 IU, ve daha sonra 3 ile tüp yıkayın - - 300 enjekte serum fizyolojik 5 ml. </li></ol></li><li> Bunun ardından, verici tracheotomize: <ol><li> 22-bıçaklı neşter kullanılarak juguler çentik ve mandibulanın arasındaki yumuşak dokularda bir orta hat kesi olun. Metzen kullanarak orta hatta tiroid bezinin kapsülü NüfuzKünt diseksiyon kullanarak baum makas, ve ayrı onun loblar. </li><li> Künt diseksiyon kullanarak, trakea ön yüzeyini açığa orta hatta boyun askısı kasları ayırın. </li><li> Açık açık trakea etrafında bir çember uçağı incelemek için kavisli bir forseps kullanın. 4-0 ipek kravat ile trakea kuşatmak. </li><li> Iris makas kullanarak, tiroid kıkırdak yaklaşık 5mm aşağı, ön trakea içinde enine bir kesi yapmak. Yavaşça trakeal kanül (14 G IV) tanıtmak ve 4-0 ipek kravat kullanarak yerine sabitleyin. </li><li> Mekanik ventilatöre trakeal kanül bağlayın. Ventilatör devresi aracılığıyla oksijen ve izofluran akışını Redirect ve hızı ve ağırlığı 13 tarafından tahmin tidal hacimde donör havalandırın. </li></ol></li><li> Xipisternum altına juguler çentik at kesi uzanan, (22 bıçaklı neşter kullanılarak) göğüs orta hatta bir kesi yapmak. </li><li> Orta hatta kalıyorumBir kemik kesici kullanılarak medyan sternotomi gerçekleştirin. Kendinden istinat retraktörün ile sternum kenarlarını geri çekin. Perikard ve plevra boşlukları girin. </li><li> Bir timektomi gerçekleştirin. İlk açık açık orta hatta timus bölmek ve sonra künt ve keskin diseksiyon bir arada kullanarak çevre yapılardan ayırmak için en kolay yoldur. NOT: timüs uzakta üstün sternum kenarlardan diseksiyon sırasında internal torasik arterlerin kökeni yaralı olabilir. Kanamayı önlemek için, hemostatik klipler bu noktalarda timus çıkarmadan önce tatbik edilebilir. </li><li> Metzenbaum makas kullanarak ve / veya keskin Lauer, inferior vena kava (IVC) uzak yağ peri-kaval teşrih. IVC altta diyaframa, superior cavo-atriyal kavşağına yağ ve bağ dokusu görece özgür olduğundan emin olun. </li><li> Keskin Lauer, çevresel ücretsiz superior vena kava (SVK) kullanma ve 4-0 ipek kravat ile kuşatmak. </li><li> Sonraki, teşrihçevredeki yapılardan ücretsiz vena kava sol ve proksimal ve distalde 4-0 ipek kravatlar ile bağlanır. Sol subklavian arteri ortaya çıkarmak için cava müdahale kısmını rezeke. </li><li> Çevresel ücretsiz keskin Lauer kullanarak arkus aorta gemiler. Innominate artere yakın ve uzak cerrahi klipleri uygulayın ve kliplerin arasına bölün. Sol karotis arter ve sol subklavyen arter un-kırpılmış bırakın. </li><li> Sonra, 24 G IV kateter ile SVC cannulate. Önceden yerleştirilen 4-0 ipek kravat ile kateter sabitleyin. </li><li> Bir çare, ama nazik greft hasat için hazırlanın. </li><li> Keskin bir makas kullanarak, diyafragma yüzeyine IVC sadece üstün bölün. Yakındaki havayolu yapıları yaralı olmadığını sağlanması, onun kanülasyon sitesine SVC üstün bölün. </li><li> Vantilatör kapatın ve trakeal kanül ayırın. Proksimale trakea transect. </li><li> Forseps ile kesilen trakea tutun ve kalp kaldırmakve akciğerler en blok. Kalp-akciğer greft kaldırılır gibi bu keskin diseksiyon ile nazik çekiş gerektirecektir. Altta yatan özofagus trakea ayırın. Bunun büyük bir kısmı hasattan sonra bozulmadan kalır, böylece inen aort yaralanmasına kaçının. </li></ol> Kalp-akciğer Greftinin 2. Hazırlık NOT: Prosedürün bu bölümünü tamamlarken, bir asistan alıcı kemirgen anestezi ve greft implantasyonu için hazırlık olmalıdır. <ol><li> Aşağı bakacak şekilde kalbin ön yüzeyi ile cerrahi gazlı bez üzerine kalp-akciğer greft yerleştirin. Keskin diseksiyon kullanarak, arka hava yollarına yaralanmasına kaçınarak, herhangi bir kalıntı özofagus çıkarın. </li><li> Inen aorta bulun ve bir 16 G künt uçlu kanül yerleştirin. 4-0 ipek kravat ile yerine kanül sabitleyin. </li><li> Aortik ark damarları bulun ve sol subklavian arter cerrahi klip uygulayın. Un-kırpılmış sol karotis arter bırakın alt içinizleyen de-havalandırma. </li><li> 16 G aort kanül Kardiyoplejisi boru bağlayarak kardiyoplejisinin ilk dozunu. Karotis arter geçici kardiyoplejisinin yeterli teslim edilmesini sağlamak için forseps ile tıkanmıştır gerekir. Not: bağlı aortik kanül sahip bir yararı arzu edildiği gibi kardiyopleji, sürekli ve / veya kesintili olarak tatbik edilebilir olmasıdır. 45 sn - 30 5 ml bolus 15 dk - aralıklı doz için, genellikle kardiyoplejisinin her 10 teslim. </li><li> Metzenbaum makas kullanarak ve / veya keskin Lauer, uzak aortadan peri-aort yağ teşrih. Aort kendi kanulasyon sola subklavian arter (distal kemer) den yağ ve bağ dokusu görece özgür olduğundan emin olun. </li><li> Sonra, keskin bir Lauer kullanarak sol ana bronş maruz ve 4-0 ipek kravat proksimale Arter. Iris makası kullanarak dikilen sola bronş distalinden transect. Sol pulmoner bağlanarak bir sol pnömonektomi gerçekleştirin4-0 ipek kravatlar ile arter ve ven. Ligature distalinde transect ve sol akciğeri çıkarın. </li><li> Yukarıdaki gibi, sağ üst ve orta lob hariç diğer tüm akciğer lobları kaldırın. Hastaya lobektomi gerçekleştirilmesi sürecinde IVC yaralanmasına kaçının. </li><li> Aort kanül Kardiyoplejisi boru bağlayın ve alıcı hazırlanan edilirken sürekli kardiyoplejisine yönetmek. Steril bir kaba (örneğin, bir bardak) kalp-akciğer greft yerleştirin. </li></ol> 3. Alıcı Operasyonu <ol><li> Yukarıda, donör operasyonu başına alıcı anestezisi. Kuruluk gelen hayvanın gözleri korumak için veteriner merhem kullanın. Pedal tutam tarafından sıkça anestezi onaylayın. </li><li> Vital bulguları ve anestezi derinliği izlemek için ücretsiz sağ ön ayakları bırakarak, donör operasyonu başına bu kez hayvan yerleştirin. </li><li> Xiphisternum gelen penise karın Tıraş. (Sonradan IV erişimi için), sol kasık küçük bir bölgesini tıraş. Uygula biroperatif yüzeylere ajan tüylerini giderildikten, yaklaşık 5 dakika bekleyin ve gazlı bez parçası ile saç kaldırmak. </li><li> Bir povidon-iyot veya chlorexidine tabanlı bir çözüm olan cerrahi siteleri hazırlayın. % 0.5 lidokain subkutan - 0.1 ile insizyonel siteleri sızmak. </li><li> Tracheotomize ve yukarıda belirtildiği şekilde alıcı havalandırın. Alternatif olarak, burun konisi anestezi altında alıcı korumak. </li><li> Yukarıda tarif edildiği gibi femoral damara bir kateter tanıtılması. IV tüp aracılığıyla fraksiyone olmayan heparine 500 IU, ve daha sonra 3 ile tüp yıkayın - - 300 enjekte serum fizyolojik 5 ml. </li><li> Penise xiphisternum bir 22-bıçak neşter ile orta hat karın kesi yaparak bir laparotomi gerçekleştirin. Kendi kendine istinat Retraktörü kullanarak karın duvarı çekin. Sonra, superior ve sıçan sol tarafa bağırsakları geri çekin. Sıcak, tuzlu batırılmış gazlı bez onları sarın. </li><li> Keskin örten yeniden aracılığıyla diseksiyon ile VKİ ve abdominal aorta Açığatro-periton yağ. </li><li> Kavisli damar kelepçe mevcut ve hazır bulundurun. </li><li> Çevresel proksimale ve distale IVC ve aort özgür ve 4-0 ipek kravatlar ile kuşatmak. Bu siteler arasında boşluk 3 cm - yaklaşık 2 olmalıdır. </li><li> Dikkatle IVC ve aort hem yeterli bir bölümünün kelepçe çene yukansında yer alır sağlanması, eğri damar kelepçesini uygulanır. </li><li> Bir tuzlu dolu 1 ml şırınga bağlı 25 G iğne ile IVC ön duvarında bir kesi yapmak. Donör IVC deliğin uzunluğu maç için Potts makas ile kesi uzatın. </li><li> Kendi kabından kalp-akciğer greft çıkarın ve kardiyoplejisinin bağlantısını kesmek. Venöz anastomoz gerçekleştirmek için en uygun pozisyonda alıcının karın greft yerleştirin. NOT: kalp-akciğer greft sonuçta karın yıllardan doğru apeks işaret ile, hafifçe eğik odaklı olacak alt kadranda bıraktı. </li><li> Topuk Güvenlive ayak 9-0 naylon sütür ile anastomoz biter. Dikiş uzun koluna bağlı iğne, daha sonra bağlı olan sütür kısa bir ucunu bırakarak, her iki ucunda güvenli bir düğüm. Kardiyoplejisinin bir doz yönetme. </li><li> Moda çalışan olarak, karşıt kısa dikiş koluna dikiş hattı ve kravat tam bir buçuk. Kardiyoplejisinin bir doz yönetme. </li><li> Sütür hattının diğer yarısını tamamlayın ve aşağı kravat. Venöz anastomoz tamamlandı. Kardiyopleji bir dozunun tatbik edilmesi </li><li> Orient karın yıllardan karşı kalbin apeks işaret ile kalp-akciğer greft alt kadranda bıraktı. Venöz anastomoz kıvrılmış veya bükülmüş olmadığından emin olun. </li><li> Gerilimsiz ve un-Dirsekli konfigürasyonlarda IVC ve aort anastomoz hem bakımını alıcı aorta ulaşmak için gerekli olacak donör aort uzunluğunu değerlendirin. </li><li> Bir iris makas ile (sol subklavian arter distalinde) inen aorta bölün. </li><li> Bir incisio Yapn, bir tuzlu su ile doldurulmuş 1 ml şırınga bağlı bir 25 G iğne ile alıcı aort ön duvarında. Donör aort deliğinin uzunluğu maç için Potts makas ile kesi uzatın. </li><li> 9-0 naylon dikiş kullanılarak aort alıcıya donör aort burnunu (üstün yönü) Çapa. Ardından, aortun medial iğne geçmek ve moda çalışan anastomoz yarısını tamamlamak. </li><li> Aort anastomoz topuk, sütür hattının (dikiş yönünde bir geçiş) yönünü ters ve moda çalışan anastomoz lateral yarısını tamamlamak. </li><li> Yavaşça SVC IV kateter kaldırmak ve cerrahi klip ile SVC Arter. </li><li> De-hava un kırpılmış sol karotis arter bularak aorta. Havanın serbestçe tahliye sağlayacak bir dik bir konumda arteri açık tutmak. </li><li> Kısaca d kavisli damar kelepçe çeneleri açılır ve (karotid arter kanamaya izinE-hava) 2 - 3 sn. Damar kelepçesini yeniden uygulayın. </li><li> Karotid arter cerrahi bir klip uygulayın. Kavisli damar kelepçesini çıkarın. </li><li> Sütür hatlarında kanama olup olmadığını kontrol edin. Varsa, 9-0 naylon sütür (kanamanın şiddetine bağlı olarak) uzunluğunda bir cerrahi gazlı bez ya da tamir nazik sıkıştırma uygulayın. </li><li> Donör kalp dakika içinde yenerek devam etmesi gerekir. </li><li> Alıcının vital bulgular, anestezi derinliği yakın ilgi ve gerektiği gibi ses canlandırma yönetmek. </li><li> Ya deneysel uç noktaları bağlı hayvanın karın kapatın ve anestezi kurtarmak izin veya kardiyovasküler değerlendirme için hayvan hazırlar. NOT: greft değerlendirme örnekleri yük-bağımlı ve yük bağımsız hemodinami in vivo tedbirler, in vitro Langendorff&#39;a işlevin önlemleri ve çalışma kalp modları ve (hayatta kalma cerrahi) ekokardiyografik veya MR incelemeleri dahil. </li><li> AtAkut Terminal deneyler sonunda, hayvanlar kan kaybından ötenazi. </li></ol>

<ol>
	<li>Abbott, C. P., Lindsey, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+for+heart+transplantation+in+the+rat.">A technique for heart transplantation in the rat.</a> Arch Surg. 89 (1964), 649-652 (1964).</li><li>Ma, Y., Wang, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+2+heterotopic+heart+transplant+techniques+in+rats:+cervical+and+abdominal+heart.">Comparison of 2 heterotopic heart transplant techniques in rats: cervical and abdominal heart.</a> Exp Clin Transplant. 9 (2), 128-133 (2011).</li><li>Wiedemann, D., Boesch, F., Schneeberger, S., Kocher, A., Laufer, G., Semsroth, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graft+function+after+heterotopic+rat+heart+transplant+with+an+isolated+reperfused+working+heart:+a+methodic+consideration.">Graft function after heterotopic rat heart transplant with an isolated reperfused working heart: a methodic consideration.</a> Exp Clin Transplant. 10 (2), 154-157 (2012).</li><li>Chen, Z. H., Xia, S. S. <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+of+heterotopic+heart-lung+transplantation+in+the+rat.">The technique of heterotopic heart-lung transplantation in the rat.</a> J Tongji Med Univ. 6 (2), 67-70 (1986).</li><li>Ibrahim, M., Navaratnarajah, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+abdominal+heart+transplantation+in+rats+for+functional+studies+of+ventricular+unloading.">Heterotopic abdominal heart transplantation in rats for functional studies of ventricular unloading.</a> J Surg Res. 179 (1), e31-e39 (2013).</li><li>Asfour, B., Hare, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+new+model+of+physiologically+working+heterotopic+rat+heart+transplantation+provides+hemodynamic+performance+equivalent+to+that+of+an+orthotopic+heart.">A simple new model of physiologically working heterotopic rat heart transplantation provides hemodynamic performance equivalent to that of an orthotopic heart.</a> J Heart Lung Transplant. 18 (10), 927-936 (1999).</li><li>Figueiredo, J. -. L., Nahrendorf, M., Sosnovik, D. E., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+of+a+novel+murine+working+heart+transplant+model.">MRI of a novel murine working heart transplant model.</a> Circ Heart Fail. 2 (3), 272-274 (2009).</li><li>Wen, P., Wang, X., 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=A+simple+technique+for+a+new+working+heterotopic+heart+transplantation+model+in+rats.">A simple technique for a new working heterotopic heart transplantation model in rats.</a> Transplant Proc. 45 (6), 2522-2526 (2013).</li><li>Didi&#233;, M., Biermann, D., 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=Preservation+of+left+ventricular+function+and+morphology+in+volume-loaded+versus+volume-unloaded+heterotopic+heart+transplants.">Preservation of left ventricular function and morphology in volume-loaded versus volume-unloaded heterotopic heart transplants.</a> A.Am J Physiol Heart Circ Physiol. 305 (4), H533-H541 (2013).</li><li>Gali&#241;anes, M., Zhai, X., Hearse, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+load+on+atrophy,+myosin+isoform+shifts+and+contractile+function:+studies+in+a+novel+rat+heart+transplant+preparation.">The effect of load on atrophy, myosin isoform shifts and contractile function: studies in a novel rat heart transplant preparation.</a> J Mol Cell Cardiol. 27 (1), 407-417 (1995).</li><li>Yokoyama, H., Ohmi, M., Murata, S., Nakame, T., Tabayashi, K., Mohri, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proposal+of+a+working+left+heart+model+with+a+heterotopic+transplantation+technique+in+rats.">Proposal of a working left heart model with a heterotopic transplantation technique in rats.</a> J Heart Lung Transplant. 14 (4), 706-712 (1995).</li><li>Maruyama, T., Swartz, M. T., McBride, L. R., Pennington, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Working+heart+model+of+heterotopic+heart-lung+transplantation+in+rats.">Working heart model of heterotopic heart-lung transplantation in rats.</a> J Thorac Cardiovasc Surg. 107 (1), 210-215 (1994).</li><li>Pacher, P., Nagayama, T., Mukhopadhyay, P., B&#225;tkai, S., Kass, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cardiac+function+using+pressure-volume+conductance+catheter+technique+in+mice+and+rats.">Measurement of cardiac function using pressure-volume conductance catheter technique in mice and rats.</a> Nat Protoc. 3 (9), 1422-1434 (2008).</li><li>Jespersen, B., Knupp, L., Northcott, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Femoral+arterial+and+venous+catheterization+for+blood+sampling,+drug+administration+and+conscious+blood+pressure+and+heart+rate+measurements.">Femoral arterial and venous catheterization for blood sampling, drug administration and conscious blood pressure and heart rate measurements.</a> J Vis Exp. (59), 3496-3410 (2012).</li><li>Habertheuer, A., Kocher, 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=Innovative,+simplified+orthotopic+lung+transplantation+in+rats).">Innovative, simplified orthotopic lung transplantation in rats).</a> J Surg Res. 185 (1), 419-425 (2013).</li></ol>

Yazarlar ifşa hiçbir şey yok.

Burada anlatılan teknikle Başarı çeşitli faktörlere esas alınacaktır. Aralarında Key üniforma greft soğutma ile tam kardioplejik tutuklama, toplam iskemik süresini en aza indirmek sağlanması, güvenli ve minimal kan kaybı ile ilişkili olan titiz operatif teknik benimseyen, hem donör ve alıcı hayvanların istikrarın sağlanması ve yeterli de-havalandırma greft olacaktır. Yukarıda kabul gibi, tekniğin teknik karmaşıklığı onun baş sınırlamadır. Biz birçok bak?...

The first rodent model of heterotopic abdominal heart transplantation (HAHT) was described by Abbott and colleagues in 19641. This technique, and subsequent modifications have been widely applied to characterize transplant graft function and immunologic status. The majority of HAHT techniques described involve a non-volume loaded heart2,3. Models of HAHT involving volume-loaded ventricles have been described, but they are frequently limited in one or more respects.
Heterotopic abdominal heart-lung transplantation (HAHLT) with a volume-loaded left ventricle (LV) has been described previously. Chen and colleagues4, and subsequently Ibrahim and colleagues5 described HAHLT with a single aorto-aortic (donor ascending to recipient abdominal aorta) anastomosis. The only volume load presented to the ventricle in this circulation is the coronary venous return. Asfour and colleagues described a HAHT technique in which the lung circuit was eliminated by anastomosing donor pulmonary artery (PA) to donor left atrium (LA)6. In this circulation, venous inflow to right ventricle (RV) occurs via a donor SVC to recipient IVC anastomosis, and the subsequent LV load is ejected into the aorto-aortic anastomosis. Cardiac function was partially assessed in vivo, and also in vitro using a Langendorff rig. Figueiredo and colleagues described a HAHLT model similar to our own7, but in mice. Venous inflow to the RV occurs via donor SVC to recipient IVC anastomosis. Blood subsequently passes through the single lung circulation and LV load is ejected into the aorto-aortic anastomosis. Cardiac function in their study was assessed by magnetic resonance imaging (MRI). Wen and colleagues described a unique HAHT technique in which the LV is loaded by means of a recipient aorta to donor LA anastomosis8. The LV, therefore, fills at systemic pressures. Cardiac function, and whether LV stroke volume is ejected antegradely in their model was not assessed.
Many of the techniques referenced above involve non-physiologic LV loading conditions, including the techniques whose partial LV load is represented only by coronary venous return. On the other hand, many techniques do approach physiologic LV loading. The majority of these techniques, as with the technique of Asfour and colleagues, omit the pulmonary circulation and utilize a donor PA to donor LA anastomosis6,9. The circulation described by Galinanes and colleagues10 employs a direct recipient cava to donor LA anastomosis, omitting the pulmonary circulation and the right heart. Yokoyama and colleagues achieve the same effect by ligating the donor PA and creating an interatrial communication in the donor heart (omitting donor lung and right heart circulations)11. The circulation of Maruyama and colleagues12 involves an anastomosis between donor left PA and recipient Ao, which permits LV filling via the pulmonary circulation as a conduit, but effectively excludes the right heart.
In cases where near physiologic loading conditions were met, we advance the technique of HAHLT in 2 major respects. First, to our knowledge, the exact configuration we report has not been described in rats. It is possibly the most versatile circulation for investigators wishing to study the physiology, structure, and immunology of the transplanted heart-lung graft. Second, we describe how the function of the transplant graft can be directly characterized in vivo. For this application, pressure-volume conductance catheters can be introduced directly into the LV apex of the transplant graft, which allows for complete cardiac functional characterization. 
The technique described here can be applied to both acute and chronic studies of transplant graft function, while the functional assessment may be performed either in vivo or in vitro. We present a model in which the loading conditions can be near physiologic, however the degree of ventricular loading may be manipulated both acutely and chronically by diverting venous return towards or away from the graft. Afterload conditions can also be manipulated. Because the lung and its airway are retained in this transplant configuration, investigators can re-ventilate the donor lung acutely. Uniquely, lung re-ventilation changes the composition of blood perfusing the transplant coronary arteries. Under non-ventilated conditions, blood ejected from the donor aorta is deoxygenated, and mixes with oxygenated blood in the recipient aorta. Under acutely ventilated conditions, ejected blood becomes oxygenated. Thus, transplant graft function can be compared under ventilated and non-ventilated conditions, and also under variably loaded conditions.
The protocol below describes important modifications to previously described HAHLT donor and recipient operations. It also describes an optimal technique for protecting the transplant graft throughout the period of ischemia (time between donor explant and recipient implant). Advantages of this technique include physiologic conditions potentially approaching that of an orthotopically transplanted graft, and a wide range of investigative applications. An important limitation is its technical complexity. With adequate mentoring and practice, the advantages of this technique will likely outweigh the challenges in adopting it.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Material/Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Celsior Cardioplegic Soution</td>
			<td>Genzyme</td>
			<td></td>
			<td>The solution is kept on ice throughout the procedure. We prepare our own solution, with slight modifications.</td>
		</tr>
		<tr>
			<td>Rodent Ventilator</td>
			<td>Harvard Apparatus</td>
			<td>Model 683</td>
			<td></td>
		</tr>
		<tr>
			<td>Vital Sign Monitor</td>
			<td>Nonin</td>
			<td>Model 9847V</td>
			<td>Displays SpO2 and heart rate.</td>
		</tr>
		<tr>
			<td>IV Cannulae</td>
			<td>Jelco</td>
			<td>3063</td>
			<td>24-26G x 3/4&#34; cannulae.</td>
		</tr>
		<tr>
			<td>IV Tubing</td>
			<td>CareFusion</td>
			<td>MP9259-C</td>
			<td>Short-length connector tubing (18cm).</td>
		</tr>
		<tr>
			<td>Surgical Clips</td>
			<td>Teleflex Medical</td>
			<td>001204</td>
			<td>Horizon titanium ligating clips.</td>
		</tr>
		<tr>
			<td>Sutures</td>
			<td>Ethicon, Sharpoint</td>
			<td>LA54G, AK-0107</td>
			<td>3-0 silk reel, and 9-0 prolene suture (single-armed, DR5 needle).</td>
		</tr>
		<tr>
			<td>Surgical Instruments</td>
			<td>Not Applicable</td>
			<td>Not Applicable</td>
			<td>The instruments used are generic, and can be purchased from any surgical supply company.</td>
		</tr>
	</tbody>
</table>

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.

Yukarıda tarif edilen HAHLT tekniği son derece teknik ve ayrıntı. Tablo yakın ilgi gerektiren 1 golleri başarısız prosedürlere karşı başarılı ile ilişkili önemli faktörlerden bazıları ve teknik zorlukları gidermek için bir rehber olarak kullanılabilir. Alıcı aort kelepçelenmemiş sonra, greft koroner arterler oksijenli kan ile doldurmak için görülmelidir. Buna göre, miyokard pembe ve iyi perfüze haline gelmelidir. Teknik olarak ...

Watch this Scientific Journal Video about Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration at JoVE.com

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

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.

Sıçan Heterotopik Abdominal Kalp / Bir Ses yüklenen Yapılandırması Tek akciğer transplantasyonu

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

rat-heterotopic-abdominal-heartsingle-lung-transplantation-volume

Research

JoVE Journal

Medicine

8.3K Görüntüleme.  UBC James Hogg Research Center, St. Paul's Hospital. 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.

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

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Sıçanlarda Ortotopik Karaciğer Nakli

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Tıp

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

Heterotopik Yardımcı Sıçan Karaciğer Nakli ile Akut Karaciğer Yetmezliği Portal Ven arteriali Akış ayarlı

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

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.

Mürin Heterotopik Kalp Nakli Tekniği

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.

Heterotopik Kalp Nakli bir Fare Modeli Kullanarak Pulmoner Vana Nakli

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

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.

Farelerde Heterotopik Servikal Kalp Nakli

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

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.

Abdominal Fare Kalp Nakli Modelinde Kan Devresi Rekonstrüksiyonu

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.

Kemirgenlerde Heterotopik Kalp Kapağı Transplantasyonu için Basitleştirilmiş Bir Model

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.

Terapötiklerin Kardiyak Allogrefte Verilmesi için Domuz Heterotopik Kalp Transplantasyonu Protokolü

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

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.

Fare Servikal Heterotopik Kalp Transplantasyon Modeli için Modifiye Manşet Tekniği

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

A Heterotopic Mouse Model for Studying Laryngeal Transplantation

Laryngeal heterotopic transplantation, although a technically challenging procedure, offers more scientific analysis and cost benefits compared to other animal models. Although first described by Shipchandler et al. in 2009, this technique is not widely used, possibly due to the difficulties in learning the microsurgical technique and time required to master it. This paper describes the surgical steps in detail, as well as potential pitfalls to avoid, in order to encourage effective use of this technique.
In this model, the bilateral carotid arteries of the donor larynx are anastomosed to the recipient carotid artery and external jugular vein, allowing for blood flow through the graft. Blood flow can be confirmed intraoperatively by the visualization of blood filling in the graft bilateral carotid arteries, reddening of the thyroid glands of the graft, and bleeding from micro vessels in the graft. The crucial elements for success include delicate preservation of the graft vessels, making the correct size arteriotomy and venotomy, and using the appropriate number of sutures on the arterial-arterial and arterial-venous anastomoses to secure vessels without leakage and prevent occlusion. 
Anyone can become proficient in this model with sufficient training and perform the procedure in approximately 3 h. If performed successfully, this model allows for immunologic studies to be performed with ease and at low cost.

The aim of this manuscript is to describe the microsurgical steps required to perform a heterotopic laryngeal transplant in mice. The advantages of this mouse model compared to other animal models of laryngeal transplantation are its cost-effectiveness and the availability of immunologic assays and data.

Laringeal Transplantasyonu İncelemek için Heterotopik Bir Fare Modeli

Laryngeal heterotopic transplantation, although a technically challenging procedure, offers more scientific analysis and cost benefits compared to other ...