Name: murine cervical heart transplantation model using a modified cuff technique
Uploaded: 2014-10-12T14:00:00
Description: Watch this Scientific Journal Video about Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique at JoVE.com

The authors declare that they have no competing financial interests.

Animals are housed in a barrier pathogen free facility. All animals receive human care in compliance with the &#8220;Principals of Laboratory Animal Care&#8221; formulated by the National Society for Medical Research&#8221; and the &#8220;Guide for the Care and Use of Laboratory Animals&#8221; prepared by the National Academy of Sciences and published by the National Institutes of Health (NIH Publication no. 86-23, revised 1985). All experiments are approved by the Austrian Ministry of Education, Science and Culture....

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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immune+mechanisms+of+cardiac+allograft+vasculopathy+an+update.">Immune mechanisms of cardiac allograft vasculopathy an update.</a> Transpl Immunol. 1, 237-249 (1993).</li><li>Libby, P., Pober, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+rejection.">Chronic rejection.</a> Immunity. 14, 387-397 (2001).</li><li>Kouwenhoven, E. A., de Bruin, R. W., Heemann, U. W., Marquet, R. L., Ijzermans, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Late+graft+dysfunction+after+prolonged+cold+ischemia+of+the+donor+kidney+inhibition+by+cyclosporine.">Late graft dysfunction after prolonged cold ischemia of the donor kidney inhibition by cyclosporine.</a> Transplantation. 68, 1004-1010 (1999).</li><li>Kouwenhoven, E. A., Ijzermans, J. N., de Bruin, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiology+and+pathophysiology+of+chronic+transplant+dysfunction.">Etiology and pathophysiology of chronic transplant dysfunction.</a> Transpl Int. 13, 385-401 (2000).</li><li>Chen, Z. H. <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+of+cervical+heterotopic+heart+transplantation+in+mice.">A technique of cervical heterotopic heart transplantation in mice.</a> Transplantation. 52, 1099-1101 (1991).</li><li>Huang, X., Chen, D., Chen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=[A+modified+model+of+cervical+heterotopic+cardiac+transplantation+for+chronic+rejection+research].">[A modified model of cervical heterotopic cardiac transplantation for chronic rejection research].</a> Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 22, 1508-1510 (2008).</li><li>Hasegawa, T., Visovatti, S. H., Hyman, M. C., Hayasaki, T., Pinsky, 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=Heterotopic+vascularized+murine+cardiac+transplantation+to+study+graft+arteriopathy.">Heterotopic vascularized murine cardiac transplantation to study graft arteriopathy.</a> Nat Protoc. 2, 471-480 (2007).</li><li>Wang, C. Y., 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=Suppression+of+murine+cardiac+allograft+arteriopathy+by+long-term+blockade+of+CD40-CD154+interactions.">Suppression of murine cardiac allograft arteriopathy by long-term blockade of CD40-CD154 interactions.</a> Circulation. 105, 1609-1614 (2002).</li><li>Matsuura, A., Abe, T., Yasuura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simplified+mouse+cervical+heart+transplantation+using+a+cuff+technique.">Simplified mouse cervical heart transplantation using a cuff technique.</a> Transplantation. 51, 896-898 (1991).</li><li>Tomita, Y., 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=Improved+technique+of+heterotopic+cervical+heart+transplantation+in+mice.">Improved technique of heterotopic cervical heart transplantation in mice.</a> Transplantation. 64, 1598-1601 (1997).</li><li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant.">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Exp Clin Transplant. 8, 245-249 (2010).</li><li>Corry, R. J., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primarily+vascularized+allografts+of+hearts+in+mice.+The+role+of+H-2D,+H-2K,+and+non-H-2+antigens+in+rejection.">Primarily vascularized allografts of hearts in mice. The role of H-2D, H-2K, and non-H-2 antigens in rejection.</a> Transplantation. 16, 343-350 (1973).</li><li>Kamada, N., Calne, R. Y. <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+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage.">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47-50 (1979).</li><li>Zimmermann, F. 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=Techniques+for+orthotopic+liver+transplantation+in+the+rat+and+some+studies+of+the+immunologic+responses+to+fully+allogeneic+liver+grafts.">Techniques for orthotopic liver transplantation in the rat and some studies of the immunologic responses to fully allogeneic liver grafts.</a> Transplant Proc. 11, 571-577 (1979).</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=Donor+pretreatment+with+tetrahydrobiopterin+saves+pancreatic+isografts+from+ischemia+reperfusion+injury+in+a+mouse+model.">Donor pretreatment with tetrahydrobiopterin saves pancreatic isografts from ischemia reperfusion injury in a mouse model.</a> Am J Transplant. 10, 2231-2240 (2010).</li><li>Heron, I. <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+accessory+cervical+heart+transplantation+in+rabbits+and+rats.">A technique for accessory cervical heart transplantation in rabbits and rats.</a> Actapathologica et microbiologica Scandinavica Section A, Pathology. 79, 366-372 (1971).</li></ol>

Rejection of vascularized allografts entails a plethora of different steps which can only be insufficiently assessed using in vitro models. Phenomena like the sensitization of recipients, antigen processing in secondary lymphoid organs but also the differentiation and proliferation of immune-competent cells are better characterized in vivo. Animal models are thereby the ideal tool for translational research16. Mouse models still represent the gold standard in basic transplant and immunologic ...

Cardiac transplantation represents the therapy of choice for patients suffering from different end-stage heart diseases. Advances in surgical techniques, more effective prophylaxis of infections, and novel immunosuppressive regimens resulted in markedly improved outcome of organ transplantation6. However, long-term graft survival has not improved rigorously over the past years7. Chronic rejection, characterized by transplant arteriosclerosis continues to be a major obstacle to long-term graft survival8-11.
The model of heterotopic heart transplantation in mice provides an important and valid tool for analysis of immunological mechanism during acute as well as chronic rejection12-15.
To date the most frequent transplant model is still the abdominal mouse heart transplantation using the suture technique. The ascending aorta of the donor heart is anastomosed to the abdominal aorta and the pulmonary artery is anastomosed to the recipient&#8217;s inferior vena cava. Much of the microsurgical difficulty of the suture model is based on the small size of the vessels, which are sutured16,17.
In contrast to the suture model the heart is placed in the neck region of the recipient where the external jugular vein is anastomosed to the pulmonary artery and the common carotid artery the aorta of the donor.
The rationale of the development of the cervical heart transplantation model using the cuff technique was to have an animal model, which allows achieving high success rates with basic microsurgical skills that will allow broad application of this model. The major advantages of this method are the significantly less occurring anastomosis related complications like hemorrhages and thrombosis when compared to the suture model18.

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			<td>Micro vessel clip</td>
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			<td>B1 00396 V</td>
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			<td>Microscissor</td>
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			<td rowspan="2">Vesseldilatator</td>
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			<td>Microforceps</td>
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			<td>Dumont 11271- 30</td>
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			<td>Clipapplicator</td>
			<td>S &#38; T</td>
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			<td>Microvessel clip</td>
			<td>S &#38; T</td>
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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.

Figure 2 shows representative survival data of cardiac allografts from BALB/c donors after transplantation into fully MHC mismatched C57BL/6 recipients. Hearts are rejected on day 7 &#177; 1 after transplantation. Finger palpation of the transplanted allograft is a sensitive method to detect the time dependent course of allograft rejection. All syngeneic heart transplants performed in our series survived indefinitely (&#62;150 days).
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Watch this Scientific Journal Video about Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique at JoVE.com

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

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.

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

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