Name: a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction
Uploaded: 2018-03-19T12:30:00
Description: Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

This work was supported by grants from the Ministry of Education Innovation Team Development Program of China (No. IRT16R57), National Natural Science Foundation of China (No. 81470896), and a research fund for Young Talent Recruiting Plans of Xi'an Jiaotong University (RW).

The protocol was carried out in accordance with the Guidelines for the Care and Use of Laboratory Animals and was approved by the Committee on the Ethics of Animal Experiments of Xi&#39;an Jiaotong University, Xi&#39;an, Shaanxi Province, China.
NOTE: None of the procedures in this protocol was performed under a surgical microscope.
1. Preoperative Preparation
<ol>
	<li>The Design and Shape of the Magnetic Rings (Figure 1)
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		<li...

<ol>
	<li>Aller, M. 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=A+half+century+(1961-2011)+of+applying+microsurgery+to+experimental+liver+research.">A half century (1961-2011) of applying microsurgery to experimental liver research.</a> World journal of hepatology. 4, 199-208 (2012).</li><li>Aller, M. 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=The+value+of+microsurgery+in+liver+research.">The value of microsurgery in liver research.</a> Liver international : official journal of the International Association for the Study of the Liver. 29, 1132-1140 (2009).</li><li>Lee, S., Charters, A. C., Chandler, J. G., Orloff, M. J. <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+orthotopic+liver+transplantation+in+the+rat.">A technique for orthotopic liver transplantation in the rat.</a> Transplantation. 16, 664-669 (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>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> Journal of visualized experiments : JoVE. , (2012).</li><li>Nagai, K., Yagi, S., Uemoto, S., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+procedures+for+a+rat+model+of+partial+orthotopic+liver+transplantation+with+hepatic+arterial+reconstruction.">Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction.</a> Journal of visualized experiments : JoVE. , e4376 (2013).</li><li>Liu, X., He, C., Huang, T., Gu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+New+Technique+for+Reconstruction+of+Hepatic+Artery+during+Liver+Transplantation+in+Sprague-Dawley+Rat.">Development of a New Technique for Reconstruction of Hepatic Artery during Liver Transplantation in Sprague-Dawley Rat.</a> PloS one. 10, e0145662 (2015).</li><li>Jamshidi, R., Stephenson, J. T., Clay, J. G., Pichakron, K. O., Harrison, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnamosis:+magnetic+compression+anastomosis+with+comparison+to+suture+and+staple+techniques.">Magnamosis: magnetic compression anastomosis with comparison to suture and staple techniques.</a> Journal of pediatric surgery. 44, 222-228 (2009).</li><li>Pichakron, K. O., 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=Magnamosis+II:+Magnetic+compression+anastomosis+for+minimally+invasive+gastrojejunostomy+and+jejunojejunostomy.">Magnamosis II: Magnetic compression anastomosis for minimally invasive gastrojejunostomy and jejunojejunostomy.</a> Journal of the American College of Surgeons. 212, 42-49 (2011).</li><li>Gonzales, K. 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=Magnamosis+III:+delivery+of+a+magnetic+compression+anastomosis+device+using+minimally+invasive+endoscopic+techniques.">Magnamosis III: delivery of a magnetic compression anastomosis device using minimally invasive endoscopic techniques.</a> Journal of pediatric surgery. 47, 1291-1295 (2012).</li><li>Wall, J., 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=MAGNAMOSIS+IV:+magnetic+compression+anastomosis+for+minimally+invasive+colorectal+surgery.">MAGNAMOSIS IV: magnetic compression anastomosis for minimally invasive colorectal surgery.</a> Endoscopy. 45, 643-648 (2013).</li><li>Xue, F., 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=Choledochojejunostomy+with+an+innovative+magnetic+compressive+anastomosis:+How+to+determine+optimal+pressure?.">Choledochojejunostomy with an innovative magnetic compressive anastomosis: How to determine optimal pressure?.</a> World journal of gastroenterology. 22, 2326-2335 (2016).</li><li>Yan, X., 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=Portacaval+shunt+established+in+six+dogs+using+magnetic+compression+technique.">Portacaval shunt established in six dogs using magnetic compression technique.</a> PloS one. 8, e76873 (2013).</li><li>Dorman, R. M., Vali, K., Harmon, C. M., Zaritzky, M., Bass, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repair+of+esophageal+atresia+with+proximal+fistula+using+endoscopic+magnetic+compression+anastomosis+(magnamosis)+after+staged+lengthening.">Repair of esophageal atresia with proximal fistula using endoscopic magnetic compression anastomosis (magnamosis) after staged lengthening.</a> Pediatric surgery international. 32, 525-528 (2016).</li><li>Russell, K. W., Rollins, M. D., Feola, G. P., Scaife, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnamosis:+a+novel+technique+for+the+management+of+rectal+atresia.">Magnamosis: a novel technique for the management of rectal atresia.</a> BMJ case reports. , (2014).</li><li>Shi, 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=Magnetic+ring+anastomosis+of+suprahepatic+vena+cava:+novel+technique+for+liver+transplantation+in+rat.">Magnetic ring anastomosis of suprahepatic vena cava: novel technique for liver transplantation in rat.</a> Transplant international : official journal of the European Society for Organ Transplantation. 28, 89-94 (2015).</li></ol>

Many clinical progresses in liver transplantation can be attributed to animal studies. Rat OLT is a widely used and well accepted model in the research of organ preservation, transplant immunology, physiology and pathology. However, it is also a very complicated procedure and requires advanced microsurgical skill. Despite many improvements in the procedure of rat OLT, the SHVC anastomosis remained a challenging step. In this article, we describe a novel magnetic anastomosis technique for SHVC reconstruction in rat OLT. T...

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research1,2. The first rat OLT was described by Lee et al. in 19733. In that model, all the vessels were reconstructed by the hand-suture technique. The hand-suture technique requires advanced microsurgical skills, which significantly limits its utilization. Since then, various modifications to the original protocol of rat OLT have been reported. Among them, the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) reported by Kamada et al. in 1979 is considered a major improvement to this model, as it significantly simplified the reconstruction procedures4. However, due to the short anterior wall of the recipients&#39; suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat OLT5,6,7.
The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels or other tubular structures using the attractive force between two magnets8,9,10,11. The magnetic force gradually compresses and remodels the tissue into a strong, sutureless anastomosis12,13. This compression anastomosis has been proven to be effective in humans14,15. We have designed a pair of magnetic rings specifically for the anastomosis of the SHVC in rats. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rat OLT16. The purpose of this article is to provide a detailed, step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique.

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			<td style="width:377px;">Shanghai Medical Instruments (Group) Ltd., Corp.</td>
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			<td style="width:377px;">Shanghai Medical Instruments (Group) Ltd., Corp.</td>
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			<td style="width:377px;">Shanghai Medical Instruments (Group) Ltd., Corp.</td>
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			<td style="width:377px;">Shanghai Medical Instruments (Group) Ltd., Corp.</td>
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			<td style="width:377px;">Shanghai Medical Instruments (Group) Ltd., Corp.</td>
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a rat model of orthotopic liver transplantation using a novel magnetic anastomosis technique for suprahepatic vena cava reconstruction

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep learning curve. The introduction of the cuff technique for anastomosis of the portal vein (PV) and infrahepatic vena cava (IHVC) has significantly simplified the transplant procedure in rats. However, due to the short anterior wall of the recipients' suprahepatic vena cava (SHVC), the cuff technique is very difficult to use for the reconstruction of the SHVC. Most researchers in this field still use the hand-suture technique for SHVC reconstruction, which makes it the bottleneck step in rat orthotopic liver transplantation. The magnetic anastomosis technique (i.e., magnamosis) is a method of connecting two vessels using the attractive force between two magnets. Our recent study has shown that the magnetic anastomosis technique is superior to the hand-suture technique for SHVC reconstruction in rats. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using the novel magnetic anastomosis technique. In this model, the reconstruction of the PV and IHVC was performed by the standard cuff technique, while the reconstruction of the bile duct (BD) was performed by a stent technique. The hepatic re-arterialization was not performed. The magnetic anastomosis technique made SHVC reconstruction much easier and significantly shortened the anphepatic phase. After a reasonable learning curve, even researchers without advanced microsurgical skills can produce reliable and reproducible results using this rat model of OLT.

After approximately 10 attempts, the magnetic anastomosis technique for SHVC reconstruction was successfully mastered by a researcher who had no prior microsurgical training. The reconstruction of the SHVC took less than 2 min. The anhepatic phase for the recipient rats was approximately 10 min. No thrombosis, bleeding or angiostegnosis was observed at the SHVC anastomotic site at 1, 5 and 30 days after reperfusion. The inferior vena cavography was performed at 2 weeks after transplantati...

Watch this Scientific Journal Video about A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction at JoVE.com

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

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