Name: orthotopic aortic transplantation in mice for the study of vascular disease
Uploaded: 2012-11-28T12:00:00
Description: Watch this Scientific Journal Video about Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease at JoVE.com

This work was funded by the core resource of the NIH P30 O'Brien center (DK 079337).

1. Presurgical Preparation
<ol>
	<li>Surgical procedures, no matter how carefully done, result in significant stress. To minimize the effects of such stress and to maximize reserves, animals should be maintained in the vivarium for at least 72 hr before use10,11.</li>
	<li>Surgical tools, gauze, and swabs must be sterile. It is not necessary to use sterile gloves providing only the tips of the instruments touch suture or the operative field.</li>
	<li>For hemostasis, it is imperative that clamping pressures ...

<ol>
	<li>Sun, H., 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+surgical+technique+for+the+establishment+of+a+murine+model+of+aortic+transplantation.">Improved surgical technique for the establishment of a murine model of aortic transplantation.</a> Microsurgery. 18, 368-371 (1998).</li><li>Dambrin, C., Calise, D., Pieraggi, M. T., Thiers, J. C., Thomsen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+aortic+transplantation+in+mice:+a+new+model+of+allograft+arteriosclerosis.">Orthotopic aortic transplantation in mice: a new model of allograft arteriosclerosis.</a> J. Heart Lung Transpl. 18, 946-951 (1999).</li><li>Calise, 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=Orthotopic+aortic+transplantation+in+rodents+by+the+sleeve+technique:+a+model+system+for+the+study+of+graft+vascular+disease.">Orthotopic aortic transplantation in rodents by the sleeve technique: a model system for the study of graft vascular disease.</a> Transpl. Proc. 33, 2369-2370 (2001).</li><li>Thomsen, 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=An+orthotopic+aortic+graft+mouse+model+to+study+the+immunopathology+of+chronic+vascular+rejection.">An orthotopic aortic graft mouse model to study the immunopathology of chronic vascular rejection.</a> Transpl. Proc. 34, 2833-2835 (2002).</li><li>Benza, R. L., George, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aortic+graft+transplantation+in+mice.">Aortic graft transplantation in mice.</a> J. Heart Lung Transpl. 21, 1319-1321 (2002).</li><li>Chereshnev, I., 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=Mouse+model+of+heterotopic+aortic+arch+transplantation.">Mouse model of heterotopic aortic arch transplantation.</a> The Journal of surgical research. 111, 171-176 (2003).</li><li>Cho, H. 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=Improved+surgical+technique+for+heterotopic+aortic+transplantation+in+mice.">Improved surgical technique for heterotopic aortic transplantation in mice.</a> J. Korean Med. Sci. 22, 12-15 (2007).</li><li>Zhong, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ+transplantation+in+mice:+current+status+and+future+prospects.">Organ transplantation in mice: current status and future prospects.</a> Microsurgery. 19, 52-55 (1999).</li><li>George, J. F., Pinderski, L. J., Litovsky, S., Kirklin, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Of+mice+and+men:+mouse+models+and+the+molecular+mechanisms+of+post-transplant+coronary+artery+disease.">Of mice and men: mouse models and the molecular mechanisms of post-transplant coronary artery disease.</a> J. Heart Lung Transpl. 24, 2003-2014 (2005).</li><li>Gorska, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+in+laboratory+animal+research+for+experimental+purposes.">Principles in laboratory animal research for experimental purposes.</a> Med Sci Monit. 6, 171-180 (2000).</li><li>Landi, M. S., Kreider, J. W., Lang, C. M., Bullock, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+shipping+on+the+immune+function+in+mice.">Effects of shipping on the immune function in mice.</a> Am. J. Vet. Res. 43, 1654-1657 (1982).</li><li>Sun, H., 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+surgical+technique+for+the+establishment+of+a+murine+model+of+aortic+transplantation.">Improved surgical technique for the establishment of a murine model of aortic transplantation.</a> Microsurgery. 18, 368-371 (1998).</li><li>Chen, B., 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=Carbon+monoxide+rescues+heme+oxygenase-1-deficient+mice+from+arterial+thrombosis+in+allogeneic+aortic+transplantation.">Carbon monoxide rescues heme oxygenase-1-deficient mice from arterial thrombosis in allogeneic aortic transplantation.</a> Am. J. Pathol. 175, 422-429 (2009).</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=Heart+transplantation+in+congenic+strains+of+mice.+Transpl.">Heart transplantation in congenic strains of mice. Transpl.</a> Proc. 5, 733-735 (1973).</li><li>Chang, E. I., 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=Vascular+anastomosis+using+controlled+phase+transitions+in+poloxamer+gels.">Vascular anastomosis using controlled phase transitions in poloxamer gels.</a> Nature medicine. 17, 1147-1152 (2011).</li></ol>

Mouse models of aortic transplantation provide a number of advantages because mice are very well defined immunogenetically9,12,13, and they can be easily manipulated to alter their expression of specific genes, if desired. As noted in the introduction, vascular surgery in the mouse is more difficult than most models because of the size of the vessels. Even the great arteries, such as the aorta are usually no more than 100-200 &mu;m in inner diameter; so manipulating these vessels requires a significant amount ...

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orthotopic aortic transplantation in mice for the study of vascular disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

Figure 1 shows an aortic graft. The white arrows denote the suture lines. A patent graft will show a visible pulse. Figure 2 indicates a typical experiment in which recipient survival was followed for a period of 56 days. One group consisted of wild-type (C57BL/6 x FVB) recipient mice transplanted with BALB/c aorta. The other group, designated "KO" consists of recipients (C57BL/6 x FVB) deficient in expression of heme oxygenase-1, which results in thrombosis of the grafts within 24 hr. N...

Watch this Scientific Journal Video about Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease at JoVE.com

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

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