Name: a novel microsurgical model for heterotopic, en bloc chest wall, thymus, and heart transplantation in mice
Uploaded: 2016-01-23T14:00:00.000+00:00
Duration: 12 min 24 s
Description: 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

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

Tüm operatif prosedürler Johns Hopkins Üniversitesi&#39;nde ve ABD Tarım Bakanlığı ve Halk Sağlığı Servisi gereklerine uygun olarak tamamlanmıştır. Bu protokol Johns Hopkins Üniversitesi Hayvan Bakım ve Kullanım Komitesi izler, kurumsal inceleme kurulu (protokol numarası M013M490) yönergeleri onayladı. Nihai sağkalım verileri aşağıda açıklanan cerrahi prosedürler için kaydedildi. Hem verici hem alıcı hayvanlar 0.1 mg buprenorfin kullanılarak rüçhan anestezisi / kg sc bir saat önce cerrahi ve alıcı hayvan buprenorfinin nakli sonrasında aynı dozda uygulanan yeniden ilk 48 saat içinde gerektiğinde yeniden dozlanır ameliyat sonrası. 1. Donör Allogreft Kurtarma Not: Alıcı anestezi süresini en aza indirmek için erken alıcı nakli daha nakli 40 dakika donör bölümünü başlayın ve eşzamanlı bitiş saati veya biraz kulak kolaylaştırmak içinAlıcı hazırlık karşı lier bitiş zamanı. <ol><li> Prosedür standart steril mikrocerrahi aletleri ve steril eldiven kullanın. Laboratuvarımız mikrocerrahi aletleri otoklav sterilizasyon kullanır. </li><li> % 4 izofluran indüksiyon buharlaştırıcı kullanılarak donör fare (erkek) anestezisi. Kullanımı atravmatik mekanik makası servikal, torasik saç ve karın bölgesini kaldırın. Yatar pozisyonda hayvan yerleştirin ve bir burun konisi ile% 1-2 üzerinde izofluran korumak. Periyodik ayak tutam çekilme refleksi değerlendirerek işlem boyunca yeterli anestezi sağlayın. </li><li> Cilt kesisi öncesinde, yaygın steril pamuklu çubuk kullanarak izopropil alkol ardından antiseptik povidon iyot uygulayarak operatif hazırlar. </li><li> Servikal ve karın derisi boyunca makas ile yüzeysel enine cilt kesisi ile başlayın. Bilateral orta aksiler çizgisinde her iki kesiler bağlayın. </li><li> Mikrocerrahi forseps kullanarak6-0 ipek sütür ve makasla eksternal juguler venler, tespit bağlanır ve bölmek için bilateral servikal bölgeyi incelemek. Sonra kullanarak elektrokoter bilateral internal juguler venler ve karotis arterler maruz sternokleidomastoid kasları bölün. Sol taraflı ve toplu bir şekilde karotis ve internal juguler venler sağ taraflı altında 6-0 ipek sütür geçirin. NOT: Bunlar bağladılar ve Adım 1.9 daha sonra bölünecektir. </li><li> Keskin, servikal bölgede kalan ekleri ücretsiz makas kullanarak, trakea anterior bulunan kayış kasları ve ilişkili gevşek areolar dokuyu, bölün. </li><li> Bipolar elektrokoter ve keskin diseksiyon kullanarak, subklavian damarları ve Arter (6-0 ipek dikiş) ortaya çıkarmak ve proksimale bölmek için pektoralis majör kas ve clavicles bölün. </li><li> Sonra, yavaşça, kavramak ve hayvanın penisi çekme. Penisin sırtında boyunca penisin dorsal ven görselleştirmek ve dezenfekteizopropil alkol ile bölge. 30 G iğne kullanarak, intravenöz dorsal ven yoluyla heparin 30.000 adet enjekte ve penis tekrar eski konumuna geri tepme için izin verir. Çevre dokuya heparin solüsyonu kısmi akabilir. </li><li> Karotis ve internal juguler ven etrafında önceden yerleştirilen dökme bağları kullanarak, bilateral, bağlanır ve yapıları bölün. </li><li> Sonraki kullanım makas enine intraabdominal kesi yapmak için. Infrahepatic inferior vena kava ortaya çıkarmak ve infrahepatic inferior vena kava içine soğuk Euro-Collins kardiyopleji solüsyonuna 2 ml enjekte etmek için bağırsakları eviscerate. Önce aşağıdaki adıma ilerleyen karaciğer renk değişikliği ve kalp atışı durması görselleştirerek uygun enjeksiyon sağlayın. NOT: Euro-Collins çözümü, laboratuvarımızda hazırlanan özel reaktifler ve araçların tabloya bakınız edilir. </li><li> Kullanımı makas ikili diaph üzerinden intratorasik boşluğu erişmekmaruz karından kesi ragmatic. Interkostal kaslar ve kaburga ile sefalad kesi uzatın. Aynı anda göğüs duvarı boyunca internal torasik damarların korunmasını sağlarken kalp, timus ve büyük damarları açığa göğüs duvarı düşünün. </li><li> Soğuk Euro-Collins kardiyopleji solüsyonuna 4 ml suprahepatik inferior vena kava enjekte edilir. </li><li> Aort kökü tespit ve inen aort distal iz. Keskin azalan aorta (maksimal uzunluğu koruyarak) kesti. </li><li> Pulmoner gövde belirlemek ve (maksimal uzunluğu koruyarak) şube noktasına sadece yakın bölün. Sonra soğuk Euro-Collins kardiyopleji solüsyonuna 2 ml kullanılarak, akciğer gövde lümenine bir yumuşak plastik uç kateter yerleştirerek pulmoner gövde ve kalp yıkayın. </li><li> 6-0 ipek sütür, ligate kullanarak ve pulmoner venlerin izdiham inferior vena kava bölmek ve bilateral superior vena kava aksesuar dalları. Sonra yükseltmesineve hava yolunu girmek için özenle ana kök bronşlarda ve trakea boyunca ekleri kalp sefalad teşrih. Keskin ve bipolar elektrokoter kullanılarak göğüs duvarı, timus ve kalp tamamen donör fare onu kurtarmanın incelemek. </li><li> Son olarak, sternum ve yanal costadan boyunca, makas kullanılarak, daha küçük boyuta allogreft göğüs duvarı ex vivo süs özenle internal torasik damarlar (Şekil 1A) bozmak için değil. Revaskülarizasyon aşağıdaki kanamayı en aza indirmek için, osteomusculocutaneous sternum sınırları boyunca bipolar elektrokoterizasyonunun kullanın. </li><li> Alıcı içerlek hazır değilse Euro-Collins solüsyonu soğuk (4 o Celcius) 10 ml allogreft yerleştirin. Alıcı içerlek için hazır olup olmadığını ancak, düz alıcı ameliyat alanına allogreft aktarın. </li></ol> 2. Alıcı Hazırlık Not: Alıcı anestezi süresini en aza indirmek için,Ayrı bir ameliyat istasyonunda önceki donör allogreft hasat tamamlanması yaklaşık 40 dakika alıcı hazırlık başlar. <ol><li> Standart steril mikrocerrahi aletleri ve prosedür için steril eldiven ayrı bir kümesi kullanın. </li><li> % 4 izofluran indüksiyon buharlaştırıcı kullanılarak (erkek veya kadın) alıcı fare anestezisi. Atravmatik mekanik makası kullanarak doğru servikal ve torasik bölgeden saç kaldırmak. </li><li> Yatar pozisyona ve açı sağ üst ekstremite hafifçe inferiora baş ve sağ üst ekstremite arasındaki 110 derecelik açı oluşturarak fare yerleştirin. Bir burun konisi ile% 1-2 izofluran üzerinde anestezi koruyun. </li><li> Bir pamuk uçlu aplikatör kullanarak fare gözleri petrol oftalmik merhem yerleştirin. Cilt kesisi öncesinde yaygın izopropil alkol ardından antiseptik povidon iyot kullanarak ameliyat sitesi hazırlamak. </li><li> Makas kullanarak, sağ alt bor boyunca orta hattan bir cilt kesi yapmakMandibula der ve sağ göğüs bölgesine infero-lateral kesi uzatmak. Mikrovasküler forseps ile künt diseksiyon kullanarak, yumuşak doku ve adventisyanın gelen çevresel serbest geminin dış juguler ven seferber. Elektrokoter kullanarak tüm şubeleri bölün ve allogreft için boş alan keskin diseksiyon ve elektrokoter kullanılarak submandibular bezin sağ lobunu çıkarın. </li><li> Bir manşet üzerine tersine çevrilmesi için eksternal juguler venin yeterli uzunluğa sahip ve 6-0 ipek sütür kullanılarak damar dış şahdamarını Arter. Bir hazır kesilmiş polimid manşet lümen yoluyla damarını yerleştirin ve yerine gemi-kaf kompleksi düzeltmek için bir buldog mikrovasküler kelepçe kullanın. Sonra proksimale evert manşet üzerine, eksternal juguler ven bölmek ve 10-0 naylon sütür ile yerde düzeltmek, makas kullanılarak. (Şekil 1B) </li><li> Karotis arter ortaya çıkarmak için bipolar elektrokoter ile sağ sternokleidomastoid kas bölün. Çevresel mafyaServikal bölgedeki uzak noktaya en sefalad arter ilize. Bu yumuşak doku ve çevresindeki adventisya kaldırmak için forseps ile kabın künt diseksiyon kullanılarak gerçekleştirilir. </li><li> 6-0 ipek sütür, ligate kullanarak ve karotis arter bölün. Bir hazır kesilmiş polimid manşet lümen yoluyla arter geçirin ve mümkün olduğunca göğüs girişine yakın bir bulldog mikrovasküler kelepçe ile yerine sabitleyin. Hafifçe evert manşet üzerine, bir mikrocerrahi dilatörü kullanarak gemiyi dilate, distale gemi bölün ve 10-0 naylon sütür ile yerde sabitleyin. (Şekil 1B) NOT: Belirli bir mikrocerrahi dilatör özel reaktifler ve araçların tabloda açıklanmaktadır. </li></ol> 3. Allogreft Ankastre <ol><li> Bir baş aşağı ve eğik pozisyonda alıcı servikal bölgede allogreft yerleştirmek için standart steril enstrümantasyon ve steril eldiven koruyun. </li><li> Daha sonra, verici inen AOR yerAlıcının arteriyel manşet yapısının üzerinde tik lümen ve 10-0 naylon sütür (Şekil 1C ve 1D) ile yerine onu bunu düzeltin. </li><li> Donör pulmoner arter ve alıcı fare (Şekil 1C ve 1D) everte eksternal juguler ven-kaf yapı arasındaki adım 3.2 ile aynı anastomoz moda. </li><li> İlk venöz mikrovasküler kelepçesini (eksternal juguler ven kelepçe) kaldırmak ve daha sonra arteriyel kelepçesini (karotis arter kelepçesi) bırakın. Arteriyel reperfüzyon sırasında, herhangi bir kanama ele allogreftin bütününü kontrol edin. Kanama görüntülenmiştir ise, kan kaybını en aza indirmek ve bipolar elektrokoter kullanılarak kanama kaynağını azaltmak için arteriyel kelepçesini yeniden uygulayın. </li><li> Greft kontrol edin ve hemostaz sağlamak. Bırakın ve tamamen arter mikrovasküler kelepçesini çıkarın. Kalp boşluklarının hızlı hacim genişlemesi ile anında görülecektir reperfüzyon belirtileri göstermeye kalbi gözlemleyin, ve b bekleyin0.5-1 dakika içinde başlayacak yeme. Kalbi nemlendirmek için ılık tuzlu su (35 ° C) kullanın. </li><li> Anastomozlar üzerinde herhangi bir dolaşması ya da tansiyonu teşvik etmeyecek şekilde anatomik pozisyona göğüs duvarı örtün. 6-0, sürekli naylon dikiş (Şekil 1E) kullanılarak cerrahi yaranın cilt kapatın. </li></ol> 4. Ameliyat Sonrası Bakım <ol><li> Sıvı replasmanı için hemen ameliyat sonrası bir 0.3 ml normal tuzlu su intraperitoneal sıvı bolus yönetin. </li><li> Daha sonra deri altından, sırasıyla, ağrı ve enfeksiyon profilaksisi için buprenorfin (0.1 mg / kg) ve enrofloksasin (5 mg / kg) enjekte edilir. </li><li> Anestezi uyanan kadar bir ısı lambası altında hayvan yerleştirin ve sternum yatma dönün. Kurtarma sırasında, yeterli allogreft perfüzyon sağlanması allogreftin fibrilasyon kalp görselleştirmek için boyun inceleyin. </li><li> Bir kez uyanık ve yatar pozisyonda, diğer farelerin şirket olmadan ayrı bir kafes (fare iade) O yiyecek ve su ad libitum alabilirsiniz nerede. Sağ üst ekstremite geçici minör kısıtlayıcı harekete, kafesin katta bir jelatin gıda kaynağı bırakın. </li><li> Ameliyat sonrası 1 saat alıcı fare gözlemleyin ve daha sonra gıda ve su ad libitum alabilir ve aktivite ve beslenme alımı için ilk 24 saat günde üç kez denetlenir kafes tesisine iade edin. İlk 72 saat için gereken günde iki defa deri altından ağrı ve sıkıntı ve buprenorfin (0.1 mg / kg) ile yeniden dozunun belirtileri için fareler izleyin. Daha sonra günlük hayvanları inceler ve her hafta onları tartın. </li><li> Herhangi fareler ağrı, sıkıntı belirtileri gösterir, ve yem tüketiminde azalma ise bir veteriner personel ile danışın. Erken ötenazi düşünün (protokolde ötanazi tekniği sonra servikal dislokasyon yapılmıştır, 7 dakika boyunca CO2 aşırı doz kullanmaktadır). </li><li> Allogreft kalp atışının durması s olmak için fareyi isteyen belirli bir son nokta olarak tanımlanıracrificed. </li></ol>

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

Dahil allotransplantasyon, immunolojik soruşturma içine faktör ama akut ve kronik rejeksiyon, doğrudan ve dolaylı antijen sunumu, alıcı duyarlılık ya da karışık kimerizmin indüksiyon mekanizmaları bunlarla sınırlı değildir fenomen çok sayıda bulunmaktadır. 19 hayvan modelleri haline gelmiştir transplant immünolojinin çalışma için altın standardı ve fare modelleri halk nedeniyle düşük bağıl veterinerlik ve konut taleplerini azalma maliyet transjenik ve gen knockout farelerde m...

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><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 500 ml solution add the ingredient listed below to a 330 ml of double distilled water. Mix well, and then fill in the rest of the 170 ml of double distilled water into the solution to a final volume of 500 ml.&lt;br/&gt; Ingredients: 1.02 g KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;, 3.66 g K&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt;, 0.56 g KCl, 0.42 g NaHCO&lt;sub&gt;3&lt;/sub&gt;, and 17.52 g of glucose.</td></tr><tr><td>Suture</td><td>Ethilon</td><td>MWI 72667</td><td>6-0 Ethilon


https://www.mwivet.com (MWI - Veterinary Supplies)</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&quot;&amp;nbsp;</td></tr><tr><td>Microsurgical dilator</td><td>S&amp;amp;T</td><td>D-5a.1</td><td>Dilator, 11 cm, FH, 0.1 mm 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.

Singenik C57BL / 6 nakli uzun süreli yaşam elde etti. Allogreft (Şekil 1) tasarım bir hayvan hayatta kalma perspektifi ve sürekli allogreft hayatta değerlendirmek yeteneği başarılı olduğunu kanıtladı. Bu canlı, aktif devam eden allogreft saç büyümesini kalan örten deri yoluyla gösterilmiştir ve kalp atışları görselleştirme ve parmak palpasyonla değerlendirilecek başardık. Hayatta kalma verileri, ayni atadan gelme nakledilen farelerde, Şekil 2&#39;de gösteri...

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.

Farelerde A Novel Mikrocerrahi Heterotopik Modeli, En Bloku Göğüs Duvarı, Thymus ve Kalp Nakli

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 Görüntüleme.  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

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.

Farelerde Heterotopik Kalp Nakli

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

Biyoloji

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

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

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

Modifiye Manşet Tekniği Sıçangiller Servikal Kalp Nakli Modeli

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

Tıp

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.

Heterotopik Fare Kalp NAKLİ için Modifiye Yöntem

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

Murine Heterotopic Heart Transplant Technique

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

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

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 ...

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 ...

Optimization of the Cuff Technique for Murine Heart Transplantation

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

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

Murin Kalp Nakli için Manşet Tekniğinin Optimizasyonu

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

İmmünoloji ve Enfeksiyon

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.

Ortootopik kalp transplantasyonunun klinik öncesi porcine modeli

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.

Vasküler Manşetler Kullanılarak Fare Heterotopik Servikal Kardiyak Transplantasyonu

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

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

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

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

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 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 ...