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

Bütün deneyler Alman Kalp Merkezi Münih ve Bavyera, Almanya (Gz. 55.2-1-54-2532.3-17-13) Hükümeti tarafından kabul edildi. Hayvanlar belli bir patojen serbest tesis içinde yer ve Tıbbi Araştırmalar Ulusal Derneği ve Ulusal Bilimler Akademisi tarafından verilen &quot;Laboratuvar Hayvanları Bakım ve Kullanım Kılavuzu&quot; ile formüle &quot;Laboratuar Hayvan Bakımı İlkeleri&quot; ile uyumlu insan bakım alınmıştır Fen ve Ulusal Sağlık Enstitüleri (NIH Yayın no. 86-23, 1985 revize) tarafından yayınlandı. NOT: İki kavisli bir forseps ve damar dilatatörler dahil steril mikrocerrahi aletlerin tam bir set, prosedür (Şekil 2a) için gereklidir. Perioperatif antibiyotik profilaksisi rutin gerekli değildir, ancak, bağışıklığı bozulmuş hayvanlarda endike olabilir. Hayvanlar, terminal izofluran inhalasyonu ile sakrifiye edildi. Donör Hazırlama ve Aşı Hasat &lt;ol&gt; <li> Intraperitonal (ip) ile hayvan midazolam enjeksiyonu (5.0 mg / kg), medetomidin (0.5 mg / kg) ve fentanil (0.05 mg / kg vücut ağırlığı) anestezisi. Operatif sahada yatar pozisyonda fare yerleştirin ve pedal çekme refleksi cerrahi toleransı onaylayın. </li><li> Klorheksidin ile cilde üç kez dezenfekte ve orta hat karın kesisi gerçekleştirin. Inferior vena kava maruz sola ıslak bir gazlı bez ile bağırsakları geri çekin. </li><li> Antikoagülasyon inferior vena kava ile 0.4 ml serum fizyolojik içinde sulandırılmış heparin 50 IU enjekte edilir. Heparin sonrası damar 1 dakika İnsizyon. </li><li> Diyaframı açın ve kalp maruz ikili torakotomi gerçekleştirin. Ön göğüs duvarı superiora yansıyan ve bir kelepçe ile yerinde tutulur. </li><li> Timus çıkarın ve açıkça aort ve pulmoner gövdede arasındaki bağ dokusunu incelemek. Sağ superior vena kava İnsizyon. </li><li> Düzeyinde harekete çıkan aort delinme30 G iğne ile brakiyosefalik gövde 5 ml şırınga üzerine monte ve yavaş yavaş 4 ° C soğuk kardiyoplejik çözeltisi 3 ml aşırı baskı olmadan retrogradually kalbi serpmek. Koroner dolaşım içine hava girmesini önleyin. Ayrıca kardiyak arrest geliştirmek için greft üzerine buz tuz çözeltisi bırakın. </li><li> Brakiosefalik gövde kökeni proksimale aort transect. </li><li> 8-0 ipek ile kalbe yakın alt ve üst venae hem kava inferior bağlanır ve bitişik harfler distal onları transect. </li><li> Kadar distale mümkün olduğunca pulmoner gövde ücretsiz teşrih. 8-0 ipek ile çatallanma yakın pulmoner arterleri bağlanır ve distalde onları transect. </li><li> Tek bir 8-0 ipek ligatür ile hep birlikte pulmoner venlerin bağlanır ve onları transect. </li><li> Donörden kalbini çıkarıp implantasyon kadar buz gibi soğuk kardiyoplejik solüsyon içinde saklayın. </li></ol> 2. Alıcı Hazırlık <ol><li> HazırlamakBir No. 11 neşter veya mikroskop altında bir makas ile polimid boru keserek anastomozlar için manşet. Manşet, bir silindirik gövde ve damar kelepçesi yerleştirilmesi için bir uzantı (çevresinin 1/3), 1 mm (Şekil 2b) bir uzunluğu, her oluşur. </li><li> Hayvan anestezisi ve yukarıda tarif edildiği gibi, cerrahi tolerans teyit etmektedir. Sıvı replasmanı için 0.25 ml ılık tuzlu su solüsyonu subkutan (sc) enjekte edilir. </li><li> Elektrikli tıraş makinesi kullanarak sağ servikal bölgenin saç çıkarın ve sıcak bir pad üzerinde bir yatar pozisyonda hayvan yerleştirin. Solunum tehlikeye sokabilir, bu kadar ön uzuvları gerilmeden kaçınarak bant ipliklerini bacakları sabitleyin. Kuruluğunu önlemek için oftalmik merhem kullanarak gözleri örtün. Cerrahi siteyi klorheksidin ile üç kez dezenfekte edin. </li><li> Juguler çentik sağ mandibular açıdan enine deri kesi olun. </li><li> Açık açık ext seferberernal juguler ven. Iki kutuplu bir forseps ile yan dalları cauterize ve bunları transect. </li><li> Submandibuler bezden ven ile izdiham düzeyinde ligatures arasındaki dış juguler ven bölün. </li><li> Manşet harici juguler ven çekin ve manşet uzantısı üzerinde bir damar kelepçe koyarak proksimale gemi tıkamak. Bitişik harfleri çıkarın ve 01:10 heparinli serum fizyolojik ile damar lümeni sulamak. </li><li> Manşet üzerinde damar duvarını Evert ve önceden belirlenmiş bir çevresel 8-0 ipek ligatür ile sabitleyin. </li><li> Sternocleidomastoid kasının yüzeysel kısmını çıkarın. </li><li> Sağ karotis arter harekete geçirmek ve karotis bifurkasyonu altındaki ligatures arasındaki gemi transect. </li><li> Manşet ile karotis arter çekin ve manşet uzantısı üzerinde bir Yaşargil kelepçe koyarak proksimale gemi tıkamak. Bitişik harfleri çıkarın ve 01:10 heparinli serum fizyolojik ile damar lümeni sulamak. </li><li> Yavaşça dilate bir gemi dilatatörle arter lümen, manşet üzerinde damar duvarını tersine çevrilmesi ve önceden belirlenmiş bir çevresel 8-0 ipek ligatür ile sabitleyin. </li><li> Greft implantasyonu için yeterli yer kazanmak için, submandibular bezin sağ lobunu çıkarın. </li><li> Implantasyon kadar nemli ameliyat alanını tutun. </li></ol> 3. Greft İmplantasyonu <ol><li> Başaşağı medial odaklı aorta ve pulmoner arter yanal ile alıcı servikal bölgede greft yerleştirin. </li><li> Karotis arter ile manşet üzerine greft aorta çekin ve çevresel 8-0 ipek ligatür ile anastomoz sabitleyin. </li><li> Onun ön yönü üzerinde akciğer gövde insizyon ve dış juguler ven ile manşet üzerine çekin. Yine çevresel 8-0 ipek ligatür ile anastomoz sabitleyin. </li><li> Karotid arter çözülmesini aşağıdaki dış juguler ven kelepçeyi çıkarın. Kalpler kanla hemen doldurur. R artırmak içineperfusion greft üzerine 37 ° C sıcak tuzlu su çözeltisi bırakın. Normalde kalp bir dakika içinde sinüs ritmi geliştirir. Venöz yerinde manşet uzantısını kaldırın. </li><li> Doğru greft pozisyonunu sağlamak, anastomozlar torsiyon olmadan gerilimsiz ve damarları olmalıdır. Bu beri cilt koterize gemiler yara iyileşmesini bozabilir etmeyin. Kanama genellikle kendiliğinden durur. Tek bir 8-0 sürekli dikiş ile cilt kapatın. </li></ol> 4. Ameliyat Sonrası Bakım <ol><li> Sıvı replasmanı için 0.25 ml ılık tuzlu su solüsyonu sc enjekte edilir. </li><li> Narkotik nalokson (1,2 mg / kg), flumazenil (0.5 mg / kg) ve sternal yatma muhafaza edilmesi için yeterli bilinci yerine kadar sıcak bir pedi üzerinde hayvan sol sc atipamezol (2,5 mg / kg vücut ağırlığı) enjekte antagonize etmek. Herhangi bir anormal davranış kaydetmek için en az bir saat daha hayvan gözlemleyin. Tamamen geri kazanılması kadar diğer hayvanların şirkete ameliyat geçirmiş bir hayvan iade etmeyinKırmızı. </li><li> Buprenorfin ile önleyici analjezi sağlamak (0.05 mg / kg sc günde üç kez) ya da işlemden sonra 72 saat içinde minimum bir süre için yerel kurumsal protokole göre alternatif bir analjezi. </li><li> Cerrah ve bir veteriner hayvanları düzenli olarak görmelisiniz. Kilo kaybı&gt;% 15, halsizlik, şişme veya cerrahi yan enfeksiyon hayvanlar durumunda terminal izofluran inhalasyonu ile ötenazi. </li></ol>

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K., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+kidney+transplantation+in+mice:+technique+using+cuff+for+renal+vein+anastomosis.">Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis.</a> PloS one. 8, e77278 (2013).</li><li>Maglione, 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+novel+technique+for+heterotopic+vascularized+pancreas+transplantation+in+mice+to+assess+ischemia+reperfusion+injury+and+graft+pancreatitis.">A novel technique for heterotopic vascularized pancreas transplantation in mice to assess ischemia reperfusion injury and graft pancreatitis.</a> Surgery. 141, 682-689 (2007).</li><li>Sucher, 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=Orthotopic+hind-limb+transplantation+in+rats.">Orthotopic hind-limb transplantation in rats.</a> J Vis Exp : JoVE. , (2010).</li><li>Kamada, N., Calne, R. 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Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> J Vis Exp : JoVE. , (2012).</li><li>Okazaki, 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+mouse+model+of+orthotopic+vascularized+aerated+lung+transplantation.">A mouse model of orthotopic vascularized aerated lung transplantation.</a> Am J Transplant. 7, 1672-1679 (2007).</li><li>Suzuki, H., Fan, L., Wilkes, D. 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The authors have nothing to disclose.

Biz çeşitli deneysel ortamlarda% 90 üzerinde bir başarı oranı ile 200 kalp nakli tamamladık. Burada anlatılan teknik ile, tam bir prosedür, deneyimli bir cerrahın 60 dakika içinde yapılabilir. Organ ihale öncesi servikal damarları hazırlayarak toplam greft iskemi süresi 20 dakika ile sınırlı olabilir. Implantasyon (sıcak iskemi süresi) için zaman sütür tekniği 16 oranla manşet tekniği kullanılarak önemli ölçüde daha kısadır. Bunun yerine sadece da...

Kalp nakli modeli sıkça I / R-yaralanma, allogreft reddini, nakil sonrası vaskülopatisinin ve imünmodüller ajanların etkinliğini araştırmak için kullanılır. Fare modelleri bilinen immünolojik ve genetik geçmişleri, birçok kendilenmiş ve transgenik suşları ve nispeten düşük maliyetlerle deneysel durumu gibi birçok avantaj birleştirir. 1973 yılında, farelerde abdominal Kalp nakli bir teknik ilk Corry ark 1 tarafından tarif edilmiştir. Bu modelde greftler alıcının abdominal aorta ve vena kava inferior ana pulmoner arter çıkan aort uç-yan anastomoz ile revaskülarize edilmektedir. 1991 yılında, farelerde rahim kalp transplantasyonu için olan bir dikiş tekniği Chen ve ark 2 tarafından tarif edilmiştir. Bu modelde bir retrograd koroner arter perfüzyon gr çıkan aorta alıcının karotis arter anastomoz tarafından kurulanKıç. Venöz kan sağ atriuma, sağ ventrikül içine koroner sinüs yoluyla drene ve alıcı dış şahdamarından (Şekil 1) içine pulmoner arter yoluyla atılır. Karın greft implantasyonu ile karşılaştırıldığında servikal modeli daha az invaziv ve greft yüzeysel konumu ek izlem muayeneleri (örn palpasyon, ekokardiyografi, intravital floresan mikroskopi) 3,4 için çok kolay erişilebilirlik sağlar. Teknik gelişmelere 5,6 olmasına rağmen, bu modelin daha yaygın kullanımı anastomoz kaçağı ve tromboz insidansının yüksek olan küçük damarların dikilir anastomoz gerçekleştirmek için teknik zorluklar nedeniyle sınırlı kalmıştır. 1991 yılında, Matsuura ve ark. damar duvarı büyük ölçüde servikal Kalp nakli 7 modelini kolaylaştıran bir sentetik silindir, üzerinde dışa dönük olduğu kaf tekniğini tanıttı. Tekniği fu oldurther renal 8, pankreas 9, bacak 10, hepatik 11,12, 13,14 akciğer ve damar nakli 15 de dahil olmak üzere çeşitli deneysel modeller için transplant kabul etmiştir. Dikiş tekniği ile karşılaştırıldığında gereken anastomozlar için zaman ve böylece sıcak iskemi süresi kısadır ve içinde manşet tekniği kullanılarak 16 daha az vasküler komplikasyonları vardır. Ayrıca, yüksek başarı oranı ile işlemi gerçekleştirmek için mikrocerrahi az eğitim ile cerrahlar sağlar. Manşet tekniğinin bir dezavantajı serebral perfüzyon değiştirebilir karotis arter, bir önkoşul ligasyonu olduğunu. Bu videoyu yazıda farelerde servikal Kalp nakli için geliştirilmiş ve basitleştirilmiş manşet tekniği sunuyoruz.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Forceps JFC-7.2</td>
			<td>S&#38;T</td>
			<td>P-00036</td>
			<td>Curved tip 0.20 mm</td>
		</tr>
		<tr>
			<td>Vessel dilatator D-5a</td>
			<td>S&#38;T</td>
			<td>S-00124</td>
			<td></td>
		</tr>
		<tr>
			<td>Adventita scissor</td>
			<td>S&#38;T</td>
			<td>S-00102</td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular clamp B-1V</td>
			<td>S&#38;T</td>
			<td>S-00396</td>
			<td></td>
		</tr>
		<tr>
			<td>Yasargil Clip</td>
			<td>Peter Lazic</td>
			<td>65,097</td>
			<td>3.5 mm lenght</td>
		</tr>
		<tr>
			<td>Bipolar forceps</td>
			<td>Micromed</td>
			<td>148-100-011</td>
			<td>tip 0.25 mm</td>
		</tr>
		<tr>
			<td>Polyimide tubes</td>
			<td>River Tech Medical</td>
			<td></td>
			<td>19-24 gauge</td>
		</tr>
		<tr>
			<td>8.0 silk ligatures</td>
			<td>Catgut</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Custodiol HTK solution</td>
			<td>Essential Pharmaceuticals</td>
			<td></td>
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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.

Ekibimiz başarıyla yaklaşık 20 eğitim ameliyatlarından sonra açıklanan teknik gerçekleştirebilir. En zorlu kısmı kolayca geri çevirir ve çoklu girişimler duvarın yırtılmaya yol açabilir, çünkü manşet üzerinde karotis arter damar duvarını tersine çevrilmesi için olduğunu. Eğitim döneminden sonra, ortalama toplam ameliyat süresi 60 ± 8 dk idi. Yaklaşık 20 dakika donör hasat, servikal damarlar ve anastomoz ve cilt kapatılması için 15 dakika hazırlanması için 25 dakika gereklidir. <...

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

Heterotopic Cervical Heart Transplantation in Mice

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

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 ...

heterotopic-cervical-heart-transplantation-in-mice

Research

JoVE Journal

Medicine

11.0K Görüntüleme.  German Heart Center Munich, Technische Universität München. The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Video: Heterotopic Cervical Heart Transplantation in Mice

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

Farelerde Kontrollü Servikal yırtığı Sakatlık

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

Tıp

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 ...

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 ...

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 ...

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

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

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

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 ...

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.

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 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 ...