Name: Orthotopic Liver Transplantation in Rats (Video) | JoVE Journal
Uploaded: 2012-07-01T14:00:00.000+00:00
Duration: 18 min 25 s
Description: Learn orthotopic liver transplantation in rats using a revised microsurgical technique. This method improves implantation speed and accuracy for successful graft survival.

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5 minutes

CT was supported by the Swiss National Science Foundation (SCORE grant 3232230-126233). The study was supported by the Art&egrave;res Foundation, the Astellas Foundation Europe and the Boninchi Foundation.

1. Donor Operation
<ol>
 <li>Anesthetize male rats weighing 200±20 grams with isoflurane on cone mask (3% for the induction, 2% during the operation, 1 l/min air flow, FiO2 70%). </li>
 <li>Perform a large median and transverse laparotomy and cauterize the epigastric vessels on both sides. </li>
 <li>Cut the falciform ligament, and separate the left diaphragmatic vein from the suprehepatic vena cava. Finally, divide the diaphragmatic vein between two 7-0 silk ligatures. </li>
 <li>Cut the left triangular and the gastro-hepatic ligaments. </li>
 <li>Divide the hepato-esophageal ligament and artery between 7-0 ligatures (coagulation is a valid alternative). </li>
 <li>Isolate the infrahepatic vena cava down to the left renal vein. Separate the right renal vein from the surrounding tissues and divide it between a 10-0 and a 7-0 ligature, on its proximal and distal ends, respectively. </li>
 <li>Divide the right suprarenal vein between 7-0 ligatures and free the liver from its posterior ligaments by cutting under gentle traction. </li>
 <li>Isolate the gastrosplenic vein and divide it between a 10-0 and a 7-0 ligature, on its proximal and distal ends, respectively. </li>
 <li>Divide the proper hepatic artery between 7-0 ties. </li>
 <li>Isolate the duodeno-pancreatic vein and divided it between a 10-0 and a 7-0 ligature, on its proximal and distal end, respectively. </li>
 <li>Insert a 22G, 3.5 mm stent into the common bile duct by practicing a small incision at 1 cm distance from the hepatic hilum. Secure the stent in position with a 7-0 ligature. </li>
 <li>Inject 10 UI of heparin, diluted into 1 ml of normal saline, through the dorsal vein of the penis. </li>
 <li>Cannulate the vena porta (as far as possible from the hilum) with a 21G needle and gently flush the liver with 20 ml of cold ringer lactate (or other preservation solutions). At the same time, cut the vena cava below the renal veins to allow an adequate outflow. Liver cold perfusion should last between one and two minutes. </li>
 <li>Cut the vena porta below the duodeno-pancreatic vein. </li>
 <li>Cut the common bile duct distally to the ligature around the stent. </li>
 <li>Complete the incision of the infrahepatic vena cava. </li>
 <li>Cut the suprahepatic vena cava skim to the diaphragm and remove the liver. </li>
 <li>Place the liver into a basin filled with cold ringer lactate and lay the basin on an ice pad. </li>
</ol>
2. Graft Preparation
<ol>
 <li>Insert the vena porta into its cuff, then evert the vessel around the edge to expose the intima. Secure the everted segment on the cuff with a 7-0 ligature. </li>
 <li>Repeat the same operation on the infrahepatic vena cava. </li>
 <li>Position a micro-clamp (4-6 mm length) on the proximal part of the infrahepatic vena cava. This operation avoids blood loss after portal reperfusion. </li>
 <li>Overturn the liver and insert two prolene 8-0 sutures at the opposite lateral edges of the suprahepatic vena cava, from outside to inside. </li>
 <li>Store the graft (immersed into ringer lactate solution) in the refrigerator at 0-4 °C. </li>
</ol>
3. Recipient Operation
<ol>
 <li>Anesthetize male rats weighing 200±20 grams with isoflurane on cone mask (3% for the induction, 2% during the operation, 1 l/min air flow, FiO2 70%) and keep on a warm pad. Of note, recipient and donor weights should be matched (±40 grams). </li>
 <li>Inject 10 ml of normal saline subcutaneously and 0.03 g of piperacillin/tazobactam intramuscular before laparotomy. </li>
 <li>Perform a midline xipho-pubic laparotomy. </li>
 <li>Reverse the xiphoid process with an auto-static forceps in order to better expose the liver. </li>
 <li>Cut the anterior ligaments and divide the left diaphragmatic vein between two 7-0 ties. </li>
 <li>Divide the hepato-esophageal ligament and artery between 7-0 ligatures (coagulation is a valid alternative). Not shown in the video (identical to donor operation). </li>
 <li>Isolate the infrahepatic vena cava down to the right renal vein. </li>
 <li>Divide the right suprarenal vein between 7-0 ligatures and free the liver from its posterior ligaments by cutting under gentle traction (not shown in the video (identical to donor operation)). </li>
 <li>Place a 7-0 ligature around the common bile duct, just below its division. Put another ligature one millimeter below and cut in between. </li>
 <li>Isolate the hepatic artery at the hilum and divide it between 7-0 ligatures. </li>
 <li>Gently separate left and right portal branches. Pay attention not to damage the common hepatic artery or the gastro-splenic vein. </li>
 <li>Position the caval ring around the vena cava, cranially to the right renal vein. Then secure the vessel wall to the ring by placing four cardinal 9-0 sutures. </li>
 <li>Repeat the same operation with the portal ring on the vena porta. </li>
 <li>Inject 10 UI of heparin, diluted into 1 ml of normal saline, through the dorsal vein of the penis. </li>
 <li>Set the anesthesia on 0.25% and the FiO2 on 100%. </li>
 <li>Clamp the infrahepatic vena cava just above the right renal vein, then clamp the vena porta just above the gastro-splenic vein. Finally clamp the suprahepatic vena cava being sure to include at least 2 mm of diaphragm. </li>
 <li>Cut the suprahepatic vena cava close to the parenchyma. </li>
 <li>Cut the left and the right portal branches and divide the septum in between. </li>
 <li>Cut the infrahepatic vena cava close to the parenchyma, paying attention not to incise any of the sutures previously positioned. Remove the liver and place the graft into the abdominal cavity. </li>
 <li>Perform an end-to-end anastomosis between the suprahepatic vena cava of the graft and the recipient. Start with the running suture of the posterior wall from left to right, then proceed on the anterior wall from right to left. Keep the anterior suture loose until the end in order to allow a generous flushing of the interior lumen before tightening the ends. </li>
 <li>Insert the caudal branch of the approximator into the portal ring handle; then insert the vena porta of the graft into the slit of the cranial branch of the approximator, with the portal cuff finding its position into the slot. </li>
 <li>Close the approximator while flushing the recipient's vena porta with normal saline and allow the cuff to insert into the vessel. Secure the recipient's vena porta around its cuff with a 7-0 circumferential tie. </li>
 <li>Remove the approximator. Open the clamp on the suprahepatic vena cava and the vena porta in order: portal flow is now reestablished. </li>
 <li>Remove the portal ring by cutting the 9-0 ligatures with extra-fine scissors.</li>
 <li>Insert the caudal branch of the approximator into the caval ring handle; then insert the infrahepatic vena cava of the graft into the slit of the cranial branch of the approximator, with the caval cuff finding its position into the slot. </li>
 <li>Close the approximator while flushing the recipient's infrahepatic vena cava with normal saline and allow the cuff to insert into the vessel. Secure the recipient's vena cava around its cuff with a 7-0 circumferential tie. </li>
 <li>Remove the approximator. Open the microclamps on the infrahepatic vena cava of the graft and the recipient in order: venous flow is now reestablished. </li>
 <li>Perform a partial incision of the recipient's common bile duct, apply a gentle traction towards the head, and insert the free side of the stent previously inserted into the bile duct of the graft. Secure the stent with a circumferential 7-0 tie. </li>
 <li>Close the animal by layers with 5-0 sutures. Allow free water and food from wakening. </li>
 <li>Administer effective analgesia according to local institutional protocols. A minimum duration of 24 h is recommended. </li>
</ol>
Of note: end-to-end hepatic artery anastomosis can be performed, when required by the experimental design (the artery is not required for long-term survival after rat liver transplantation). 
4. Representative Results
The described technique could be successfully mastered by our team after approximately 10 training surgeries. It allowed 100% long-term (&gt;21 days) survival on 36 consecutive Lewis-to-Lewis and Dark Agouti-to-Lewis transplantations, after the short training period (a number of syngeneic liver graft recipients have been kept alive for over 12 months now). Post- syngeneic transplant day 1 liver function tests showed median aspartate transaminase (AST): 84 U/l (51.7 - 92.7) and median alanine transaminase (ALT): 172.5 U/l (127.5 - 240.2). The mean anhepatic phase (from vena porta clamping to graft re-perfusion) was 14 ±2 min, with the longest step being the suprahepatic vena cava anastomosis (running suture) time 9±2 min.
The quick-linker system allowed the positioning of 1.55 mm-bore cuffs on vena porta and 2.40 mm-bore cuffs on vena cava (recipients weighing 200 ±20 g). By contrast, when we tried transplanting using Kamada's technique, they could not exceed 1.40 mm and 2.16 mm, respectively (rats matching for strain and weight). As shown in Figure 7, quick-linker rings are designed to keep recipient's vessels on optimal stretching, allowing minimum caliper and length wasting, which results in closer-to-physiological hemodynamic results. 
<img src="/files/ftp_upload/4143/4143fig1.jpg" alt="Vascular clamp measurement; diagram with dimensions for v.cava and v.porta beside coin for scale." data-altupdatedat="1787630408000" data-alt="Figure 1"> 
Figure 1. Rings measures.
<img src="/files/ftp_upload/4143/4143fig2.jpg" alt="Microclamps and applicator tool, sizes 4mm and 6mm, for precise surgical applications." data-altupdatedat="1787630408000" data-alt="Figure 2"> 
Figure 2. Microclamps measures.
<img src="/files/ftp_upload/4143/4143fig3.jpg" alt="Surgical tools setup; includes hemostatic forceps and bone holding clamps on a textured surface." data-altupdatedat="1787630408000" data-alt="Figure 3"> 
Figure 3. Quick-linker kit.
<img src="/files/ftp_upload/4143/4143fig4.jpg" alt="Surgical suture equipment with needle holder and wire, used in medical procedures." data-altupdatedat="1787630408000" data-alt="Figure 4"> 
Figure 4. Quick-linker armed.
<img src="/files/ftp_upload/4143/4143fig5.jpg" alt="Forceps tool for precision handling, used in dissection and laboratory experiments." data-altupdatedat="1787630408000" data-alt="Figure 5"> 
Figure 5. Quick-linker closed.
<img src="/files/ftp_upload/4143/4143fig6.jpg" alt="Static equilibrium demonstration; clamp on inclined plane; physics experiment on forces and balance." data-altupdatedat="1787630408000" data-alt="Figure 6"> 
Figure 6. SHVC clamp.
<img src="/files/ftp_upload/4143/4143fig7.jpg" alt="Cuff techniques comparison diagram; vascular anastomosis; surgical process; turbulent flow illustration." data-altupdatedat="1787630408000" data-alt="Figure 7"> 
Figure 7. Classic vs. quick-linker assisted cuff-anastomosis hemodynamics.
<img src="/files/ftp_upload/4143/4143fig8.jpg" alt="Rodent skin transplantation process diagram, donor and recipient marked for surgical procedure." data-altupdatedat="1787630408000" data-alt="Figure 8"> 
Figure 8. Donor and recipient's skin preparation and incision.

<ol><li>Starzl, T. E., Marchioro, T. L., Porter, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Experimental+and+clinical+observation+after+homotransplantation+of+the+whole+liver%5BTitle%5D)+AND+%22Revue+internationale+d%27hepatologie%22%5BJournal%5D)">Experimental and clinical observation after homotransplantation of the whole liver.</a> Revue internationale d'hepatologie. 15, 1447(1965).</li>
<li>Starzl, T. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evolution+of+liver+transplantation%5BTitle%5D)+AND+%22Hepatology%22%5BJournal%5D)">Evolution of liver transplantation.</a> Hepatology. 2, 614S-636S (1982).</li>
<li>Kamada, N., Calne, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Orthotopic+liver+transplantation+in+the+rat.+Technique+using+cuff+for+portal+vein+anastomosis+and+biliary+drainage%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">Orthotopic liver transplantation in the rat. Technique using cuff for portal vein anastomosis and biliary drainage.</a> Transplantation. 28, 47(1979).</li>
<li>Kashfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat%5BTitle%5D)+AND+%22Transplantation+proceedings%22%5BJournal%5D)">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplantation proceedings. 37, 185-188 (2005).</li>
<li>Hori, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Comprehensive+and+innovative+techniques+for+liver+transplantation+in+rats%3A+A+surgical+guide%5BTitle%5D)+AND+%22World+Journal+of+Gastroenterology%3A+WJG%22%5BJournal%5D)">Comprehensive and innovative techniques for liver transplantation in rats: A surgical guide.</a> World Journal of Gastroenterology: WJG. 16, 3120(2010).</li>
<li>Oldani, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+novel+technique+for+rat+liver+transplantation+using+Quick+Linker+system%3A+a+preliminary+result%5BTitle%5D)+AND+%22Journal+of+Surgical+Research%22%5BJournal%5D)">A novel technique for rat liver transplantation using Quick Linker system: a preliminary result.</a> Journal of Surgical Research. 149, 303-309 (2008).</li>
<li>Nosaka, T., Bowers, J. L., Cay, O., Clouse, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biliary+complications+after+orthotopic+liver+transplantation+in+rats%5BTitle%5D)+AND+%22Surg.+Today%22%5BJournal%5D)">Biliary complications after orthotopic liver transplantation in rats.</a> Surg. Today. 29, 963-965 (1999).</li>
<li>Li, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High+incidence+of+biliary+complications+in+rat+liver+transplantation%3A+can+we+avoid+it%5BTitle%5D)+AND+%22World+J.+Gastroenterol%22%5BJournal%5D)">High incidence of biliary complications in rat liver transplantation: can we avoid it.</a> World J. Gastroenterol. 17, 3140-3144 (2011).</li>
<li>Peng, Y., Gong, J. P., Yan, L. N., Li, S. B., Li, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improved+two-cuff+technique+for+orthotopic+liver+transplantation+in+rat%5BTitle%5D)+AND+%22Hepatobiliary+Pancreat.+Dis.+Int%22%5BJournal%5D)">Improved two-cuff technique for orthotopic liver transplantation in rat.</a> Hepatobiliary Pancreat. Dis. Int. 3, 33-37 (2004).</li>
<li>Urata, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Decreased+survival+in+rat+liver+transplantation+with+extended+cold+preservation%3A+role+of+portal+vein+clamping+time%5BTitle%5D)+AND+%22Hepatology%22%5BJournal%5D)">Decreased survival in rat liver transplantation with extended cold preservation: role of portal vein clamping time.</a> Hepatology. 28, 366-373 (1998).</li>
</ol>

No conflicts of interest declared.

The present rat liver transplantation model can be easily established. In our experience, a few key points help minimize the risk of death (&#x2264;20%). Early deaths, occurring within the first three hours from reperfusion, are most often due to a bowel infarction, as a consequence of a long portal clamping. It is therefore recommended to keep the anhepatic phase within the ideal limit of 15 minutes (20 minutes still acceptable). Central air embolism can also be a cause of sudden death and it is compulsory to flush all veins prior to anastomosis. 
Deaths occurring between postoperative day one and three are often due to liver failure or thrombosis of the vena cava. One should therefore avoid any aggressive graft flushing before the explantation in the donor (we flush with 20 ml over approximately 60 seconds) and ensure an appropriate cold preservation (full immersion into 0-4 &deg;C ringer lactate solution). The risk of thrombosis can be decreased by minimizing the manipulation of the intima and preserving its integrity. We do not recommend the use of anticoagulation after surgery.
Deaths between postoperative day four and nine are often related to bacterial cholangitis7. Based on our experience, a single dose of piperacilline/tazobactam (0.03 g) given before laparotomy is enough to minimize the risk of common bile duct infection.
Death is a rare event after day 10. However, due to absence of arterial flow, some bile duct problems can be observed with cholastasis8. Depending on the needs of the investigator an arterialization may be considered9. Of note, a cold ischemia time up to 24 hours can be considered as acceptable10.
The quick-linker system was originally designed to perform liver transplantation with three cuff anastomoses (very short anhepatic phase). However, the three-cuff technique requires the use of very small calipers for the suprahepatic vena cava (SHVC) cuff and it has not been widely accepted due to the risk of liver outflow problems11-14. We therefore favor a quick-linker assisted three-cuff technique only when warm ischemia times shorter than ten minutes are required. In most studies however, implantation phases up to 15-18 minutes are acceptable, allowing the use of the two-cuff technique, and a more physiological SHVC drainage. As shown in this video paper, we routinely use the quick-linker system in the second part of the implantation only. While some extra surgical time is required to position the quick-linker handles, this part of the procedure is associated with minimal risks and allows for a shortening of the critical anhepatic phase thanks to easier cuffs insertions. Other advantages of the quick-linker system are a better graft-recipient vessel alignment and the use of wider cuffs with better hemodynamic results compared to the previously described techniques (including the one described by Kamada et al.)6(Fig 7).

<table><tbody><tr><td>&lt;strong&gt; Tool / Reagent&lt;/strong&gt;</td><td>&lt;strong&gt; Company&lt;/strong&gt;</td><td>&lt;strong&gt; Catalogue n.&lt;/strong&gt;</td><td>&lt;strong&gt; Comment&lt;/strong&gt;</td></tr><tr><td>Portal cuff</td><td> &lt;a href=&quot;http://altecweb.com&quot; target=&quot;_blank&quot;&gt;altecweb.com&lt;/a&gt; or &lt;a href=&quot;http://smiths-medical.com&quot; target=&quot;_blank&quot;&gt;smiths-medical.com&lt;/a&gt;</td><td> Coil 01-96-1729 800/100/420</td><td> Bore : 1.55mm &lt;br/&gt; Wall : 0.30mm &lt;br/&gt; Length: 3.5 mm</td></tr><tr><td>Infrahepatic v. cava cuff</td><td> &lt;a href=&quot;http://altecweb.com&quot; target=&quot;_blank&quot;&gt;altecweb.com&lt;/a&gt; or &lt;a href=&quot;http://smiths-medical.com&quot; target=&quot;_blank&quot;&gt;smiths-medical.com&lt;/a&gt;</td><td> Coil 01-96-1733 800/100/540</td><td> Bore : 2.40mm &lt;br/&gt; Wall : 0.30mm &lt;br/&gt; Length: 3.5 mm</td></tr><tr><td>Common bile duct stent</td><td>common 22G venflon</td><td>&amp;nbsp;</td><td>&amp;nbsp;</td></tr><tr><td>7-0 silk ties</td><td>&lt;a href=&quot;http://georgetiemann.com&quot; target=&quot;_blank&quot;&gt;georgetiemann.com&lt;/a&gt;</td><td>160-1226-7/0</td><td>&amp;nbsp;</td></tr><tr><td>10-0 prolene</td><td>&lt;a href=&quot;http://ethicon.com&quot; target=&quot;_blank&quot;&gt;ethicon.com&lt;/a&gt;</td><td>&amp;nbsp;</td><td>&amp;nbsp;</td></tr><tr><td>9-0 prolene</td><td>&lt;a href=&quot;http://ethicon.com&quot; target=&quot;_blank&quot;&gt;ethicon.com&lt;/a&gt;</td><td>&amp;nbsp;</td><td>&amp;nbsp;</td></tr><tr><td>8-0 prolene</td><td>&lt;a href=&quot;http://ethicon.com&quot; target=&quot;_blank&quot;&gt;ethicon.com&lt;/a&gt;</td><td>&amp;nbsp;</td><td>&amp;nbsp;</td></tr><tr><td>Straight micro forceps</td><td>s-and-t.net</td><td>FRC-15RM-8 </td><td>&amp;nbsp;</td></tr><tr><td>Curved micro forceps</td><td>s-and-t.net</td><td>FRS-15-RM-8 </td><td>&amp;nbsp;</td></tr><tr><td>Dissection needle-holder</td><td>s-and-t.net</td><td>B-15-8.2 </td><td>&amp;nbsp;</td></tr><tr><td>Anastomosis needle-holder</td><td>rumex.net</td><td>8-021T</td><td>&amp;nbsp;</td></tr><tr><td>Micro clamps</td><td>&lt;a href=&quot;http://finescience.com&quot; target=&quot;_blank&quot;&gt;finescience.com&lt;/a&gt;</td><td>18055-03</td><td>6 x 1mm</td></tr><tr><td>Micro clamps applicator</td><td>&lt;a href=&quot;http://finescience.com&quot; target=&quot;_blank&quot;&gt;finescience.com&lt;/a&gt;</td><td>18057-14</td><td>&amp;nbsp;</td></tr><tr><td>Suprahepatic v. cava clamp</td><td>&lt;a href=&quot;http://pharmap.ch&quot; target=&quot;_blank&quot;&gt;pharmap.ch&lt;/a&gt;</td><td>13.349.14</td><td>&amp;nbsp;</td></tr><tr><td> Quick-Linker rings</td><td> &lt;a href=&quot;http://dltchiropody.co.uk&quot; target=&quot;_blank&quot;&gt;dltchiropody.co.uk&lt;/a&gt;</td><td> PAR 10</td><td> modified scalpel blades</td></tr><tr><td> Quick-Linker approximator</td><td>pharmap.ch</td><td> UD-04-275</td><td> modified Kocher&amp;rsquo;s forcep</td></tr></tbody></table>

orthotopic liver transplantation in rats

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Orthotopic Liver Transplantation in the wrath is a demanding procedure and requires a lung microsurgical training to be mastered properly. Accuracy and speed, especially during the phase of implantation are the keys to success. In the next videos, we will reveal every fundamental step to a successful transplantation and explain how a quick clinical device can facilitate the implantation phase.Enjoy the videos. The donor is open via transverse incision. The falciform ligament is section and the inferior satic vena cava is exposed.Left diaphragmatic vein is detached from the cava, then divided between silk seven oh ties. Gas osteopathic ligament is sectioned, then esophageal ligament is divided between ties. The atic vena cava is exposed by gently pulling small en large bowel.On the left side, peritoneum is opened and the vena cava is gently freed from the surrounding tissues. The right adrenal vein is isolated and then divided between seven oh ties. Liver is pulled on the left side and gently kept under traction.Retro hepatic and right triangular ligaments are sectioned.Right. Renal vein is tied at its proximal end using a 10 oh nylon tie. Right renal vein and artery are tied together at their distal end right renal vein is then divided.Overview of the completely isolated repath vena cava is shown. Overview of the hepatic hilum and postal vein regional anatomy, proper hepatic artery, common B duct gastro splenic vein. The pancreas glu.The gastro splenic vein is isolated and divided between a 10 oh and a seven oh tie. Upper hepatic artery is sectioned between seven oh ties. Finally, the vino pancreatic vein is divided.An overview of the isolated vena porter is shown. Common bile duct is partially opened at its junction with the pancreatic tissue. A 3.5 millimeter 22 G stent is inserted and strongly secured.10 units of heparin are injected. Liver is then flashed through the vena porter with 20 milliliter of colder ized ringer lactate solution while distal vena cava is incised to allow an adequate outflow, the vena porter is cut at its distal end section of the common bowel duct is completed. F hepatic vena cava is then cut at its distal end while supra hepatic vena cava is cut, skied to the diaphragm that is removed and immersed into a four degrees Celsius Wrangler lactate solution solution.Vena porter is inserted into its calf, then diverted to expose the intima and finally secured on the calf with a seven oh tie. The same is done on the repath vena calf. A micro clamp is positioned on the very cranial part of the repath vena cava.Liver is overturned and two ATO prolene stitches are passed at the opposite lateral edges of the satic Vena cava from outside to inside Recipient is Kept under Isof. Fluor anesthesia on a warm pad, 10 milliliters. Normal cell line are injected subcutaneously before laparotomy em midline cyop pubic incision is performed and cy is retroverted in order to better expose the liver.Common bi Duct is secured with a seven oh tie and sectioned just before its bifurcation left and right. Portal branches are accurately separated. Caval ring is positioned around the vena cava cranial to the left renal vein.A first nine oh stitch is passed through the vessel wall at its right edge, then secured to the caval ring in the pre-established position. A second stitch is passed through the opposite edge and secured. The third one is secured in ventral position.Finally, the fourth one is secured in dorsal position. After having overturned the ring on the left side, the first stitch is passed through the posterior wall by lifting the vessel. The portal ring is then positioned and the tie is secured to its posterior branch.A second stitch is passed through the edge of the left portal branch and secured third one to the edge of the right portal. Branch Then secured. The last one pierces the anterior wall of the port And is secured in position Before clamping.A last cleaning of the satic vena cava is performed in repath Vena cava and vena porta are cross clamped in order. Spath vena cava is then Cross clamped and incised scheme to the parenchyma Left and right portal branches are cut and the septum in between is divided inre vena cava is cut scheme to the parenchyma. Then liver is removed.Inna cava and vena porta are safely kept opened by the two rings. Graft is positioned into the abdominal cavity and the NATO suture is secured to the left edge of the spath vena cava. A second suture is secured to the right edge.Iranian suture of the posterior wall is performed from left to right, therefore not to include the liver parenchyma into the suture. The anterior wall suture is kept loose in order to allow a generous flashing before complete closure. This maneuver avoids air embolism.The coddle branch of the approximator is inserted into the portal ring handle and the portal calf is inserted through the slit of its cranial branch. The recipient's Vina porter is flashed with normal saline while the approximator is closed. To assist the calf insertion recipient's, Porter is secured to the calf by a seven oh tie.The Approximator is removed and portal flow reestablished by removing in order the Atic VNA CVA clamp and the portal clamp portal ring is removed by cutting. The nano ligatures though is reestablished with the same technique recipients. Common bile duct is kept under traction and the stent is inserted through a small incision.A circular ligature secures its position. An overview of the fused graft is shown a.

Research

JoVE Journal

Medicine

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

We describe the design of the “quick-linker” device for easier orthotopic rat liver transplantation.

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

Auxiliary liver transplantation provides a temporary support in acute hepatic failure, until regeneration of the failing liver. The heterotopic auxiliary liver transplantation (HALT) with portal vein arterialization (PVA) renders sufficient liver function. We developed an analogous technique in the rat, to examine the influence of the portal vein arterialization on the morphology and function of the graft.

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

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.

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

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver's common hepatic artery to the recipient liver's proper hepatic artery.

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

In this protocol, we discuss the implementation of a model of successful orthotopic liver transplantation (OLT) in mice. Additionally, adjuvants to further analyze allograft patency after successful OLT in a mouse are discussed as well, specifically utilizing microcomputed tomography (microCT) scans.

Successful Orthotopic Liver Transplantation in Mice Utilizing Microcomputed Tomography Angiography

The rat heterotopic auxiliary liver transplant protocol described here offers a practical investigational tool for exploring mechanisms of hepatic allograft rejection. This model helps to alleviate the surgical hurdles and animal stress of orthotopic liver transplantation in rats.

Heterotopic Auxiliary Whole Liver Rat Transplant Model Utilizing a Hepaticoureterostomy for Allograft Rejection Studies

This protocol describes an improved technique for orthotopic left lung transplantation in rats, focusing on reducing operational difficulty and improving survival rates.

Orthotopic Left Lung Transplantation in Rats

Learn orthotopic liver transplantation in rats using a revised microsurgical technique. This method improves implantation speed and accuracy for successful graft survival.

Orthotopic Liver Transplantation in Rats

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