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)
	<ol>
		<li...

<ol>
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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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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 ...

The overall goal of this protocol is to introduce a rat orthotopical liver transplantation model using a magnetic anastomosis technique for suprahepatic vena cava reconstruction. This method can help answer key question in the liver transplantation model about suprahepatic vena cava reconstruction. The main advantage of this technique is that it reduces the anhepatic phase of liver transplantation and greatly improves the postoperative survival rate.First, shave the entire abdominal skin of the rat. Disinfect the corresponding skin with a povidone-iodine solution. Dissect the falciform ligament around the liver.Next, isolate and ligate the left inferior phrenic vein. Perform a V incision on the anterior wall. Insert the stent into the lumen of the bile duct.Make sure at least half of the stent lies outside the bile duct. Secure the stent with a 6-0 silk suture. Cut one end of the suture.Keep the other one to hold during later anastomosis. Isolate the infrahepatic vena cava down to the left renal vein level. Dissect it until the infrahepatic vena cava is skeletonized.Separate and ligate the right suprarenal vein. Separate and ligate the right renal vein. Heparinize the rat through the dorsal vein of the penis.Insert a 22-gauge catheter into the abdominal aorta below the left renal vein. Perform a thoracotomy. Clamp the thoracic aorta using a microforceps.Perfuse the donor liver through the catheter in the aorta. Transect the suprahepatic vena cava. In the meantime, cut the vena cava below the right renal veins to allow the perfusate flowing out of the liver.Dissect the connective tissue surrounding the portal vein. Ligate and divide the pyloric vein. Transect the portal vein at the level of the splenic vein.Transect the infrahepatic vena cava at the level of the left renal vein. Liver graft preparations are performed in four degree Celsius saline bath. Insert the infrahepatic vena cava into its cuff, with the cuff extension pointing towards to the liver.Evert the distal end of the infrahepatic vena cava over the cuff body. Secure the edge with a 6-0 silk suture. Repeat the above steps on the portal vein to attach a cuff to the donor's portal vein.Pull the diaphragm through the forceps, and trim the suprahepatic vena cava. Pass the suprahepatic vena cava through the magnetic ring. Evert the distal end of the suprahepatic vena cava over the ring.And secure the ring with a 6-0 silk suture. Then, remove the excess diaphragm around the liver. Place a 6-0 silk suture, and ligate the common bile duct.Isolate the common bile duct from the first hepatic portal. Ligate and divide the right adrenal vein. Place a strap in the suprahepatic vena cava of the liver.Clamp the infrahepatic vena cava just above the right renal vein with microvessel clips. Clamp the portal vein at the level of pyloric vein. Inject two milliliter normal saline through the portal vein slowly to flush the blood out of the liver.Drag down the liver by pulling the previous placed strap. Clamp the suprahepatic vena cava and part of the diaphragm with a Satinsky clamp. Excise the liver by transecting the suprahepatic vena cava just above the liver.Transect the infrahepatic vena cava close to the parenchyma and the portal vein at the porta hepatis. Insert the recipient's remaining suprahepatic vena cava into a magnetic ring. Fix the magnetic ring on a Satinsky clamp by magnetic attraction.Evert the suprahepatic vena cava over the magnetic ring. Place the donor liver graft orthotopically, and fill the lumen with normal saline solution, coupling the magnetic rings to fulfill the vessel construction. Reconstruct the portal vein.Fill the lumen of the portal vein with normal saline. Insert the cuff body of the donor's portal vein into the recipient's portal vein. And secure that with a circumferential 6-0 silk ligature.Remove the Satinsky clamp from the suprahepatic vena cava. Incise a V on the anterior wall of the common bile duct. Insert the donor stent into the lumen of the recipient bile duct.And fix it with a 6-0 silk circumferential suture. Pull the sutures on the two bile ducts together, and tie the sutures to bring the bile ducts close to each other. The angiography two weeks POD showed a patent blood flow of the suprahepatic vena cava anastomosis.The post-transplant survival rate was up to 85%to day 30 POD. The magnetic anastomosis technique makes easy and fast suprahepatic vena cava reconstruction in rats possible. After watching this video, without advanced microsurgical skills, can produce reliable and reproducible results using this rat model of liver transplantation.

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