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)

All experiments were approved by the German Heart Center Munich and the Government of Bavaria, Germany (Gz. 55.2-1-54-2532.3-17-13). Animals were housed in a specific pathogen free facility and received human care in compliance with the &#8221;Principles of Laboratory Animal Care&#8221; formulated by the National Society for Medical Research and the &#8221;Guide for the Care and Use of Laboratory Animals&#8221; provided by the National Academy of Sciences and published by the National Institutes of Health (NIH Publicatio...

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The authors have nothing to disclose.

We have completed over 200 cardiac transplants with a success rate over 90% in various experimental settings. With the technique described here, the complete procedure can be done within 60 min by an experienced surgeon. By preparing the cervical vessels before organ procurement, total graft ischemia time can be limited to 20 min. Time for implantation (warm ischemia time) is significantly shorter using the cuff technique compared to the suture technique16.
Instead of simply excisin...

The heart transplantation model is frequently used to investigate I/R-injury, allograft rejection, post-transplant vasculopathy and the efficiency of immunomodulating agents. Mouse models combine many advantages like the known immunological and genetic backgrounds, the availability of many inbred and transgenic strains and relatively low experimental costs.
In 1973, a technique of abdominal heart transplantation in mice was first described by Corry et al1. In this model grafts are revascularized by end-to-side anastomoses of the ascending aorta to the recipient&#8217;s abdominal aorta and of the main pulmonary artery to the inferior vena cava. In 1991, a suture technique for cervical heart transplantation in mice was described by Chen et al2. In this model a retrograde coronary artery perfusion is established by anastomosis of the recipient&#8217;s carotid artery to the ascending aorta of the graft. The venous blood drains via the coronary sinus into the right atrium, the right ventricle and is ejected through the pulmonary artery into the recipient external jugular vein (Figure 1). In comparison to the abdominal graft implantation the cervical model is less invasive and the superficial location of the graft enables a very easy accessibility for additional follow-up examinations (e.g. palpation, echocardiography, intravital fluorescence microscopy)3,4.
Despite technical improvements5,6, a more widespread use of this model has been limited due to technical difficulties to perform sutured anastomoses of small vessels with a high incidence of anastomosis leakage and thrombosis. In 1991, Matsuura et al. introduced the cuff technique in which the vessel wall is everted over a synthetic cylinder, which greatly facilitates the model of cervical heart transplantation7. The technique was further adopted for various experimental transplant models including renal8, pancreatic9, limb10, hepatic11,12, lung13,14 and vascular transplantation15. In comparison to the suture technique the required time for anastomoses and, thus, the warm ischemia time is shorter and there are significantly less vascular complications using the cuff technique16. Moreover, it enables surgeons with little training in microsurgery to perform the operation with a high success rate. A disadvantage of the cuff technique is the prerequisite ligation of the common carotid artery, which may alter cerebral perfusion.
In this video paper we present an improved and simplified cuff technique for cervical heart transplantation in mice.

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	<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>
			<td></td>
		</tr>
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</table>

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.

Our team could successfully perform the described technique after approximately 20 training surgeries. The most challenging part is to evert the vessel wall of the carotid artery over the cuff, because it easily flips back and multiple attempts may lead to tearing of the wall. After the training period, the mean total operative time was 60 &#177; 8 min. Around 20 min are required for donor harvest, 25 min for preparation of the cervical vessels and 15 minutes for anastomoses and skin closure.
...

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.

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

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11.0K vues.  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.