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

1. Handle: Design and Shaping
<ol>
 <li>Secure an Nr 10 carbon scalpel blade on a wooden board with a tape.</li>
 <li>Tag the center of the ring under the microscope. It should be located 1.2 cm straight from the grove of the scalpel blade.</li>
 <li>Drill a 2 mm hole with a spherical diamond bit, mounted on a high-speed drill.</li>
 <li>Widen the hole with fusiform or conic diamond bits. The final diameter varies according to the rat weight as shown in Table 1.</li>
 <li>Design the shape of the handle with a permanent marker. Figures 1 and 2 can be used as stencils.</li>
 <li>Convert the drill into a mini bench grinder and remove the part of the blade outside the shape.</li>
 <li>Round off the piece with a low speed rotating brush.</li>
</ol>
2. Approximator: Design and Shaping
<ol>
 <li>The approximator is obtained by modifying straight surgical Klemmer's or Kocher's forceps.</li>
 <li>Cut off half of one branch under the microscope using the high-speed mini bench grinder.</li>
 <li>Make symmetrical grooves on both sides to allow for the handle to slide in completely. </li>
 <li>Cut a longitudinal slit in the long arm and check that the center of the long arm and a well-positioned handle are on a same axis. </li>
 <li>Design a notch on the long arm facing the center of the handle. </li>
 <li>Make the branch as thin as possible and round off the piece with the low speed rotating brush. </li>
</ol>
3. Cuffs: Design and Shaping
<ol>
 <li>Cuff calipers vary according to the rat weight and strain. By contrast, their length should be kept around 4 mm. </li>
 <li>Take fine-bore polyethylene tubing and round off one edge with an Nr 10 scalpel. </li>
 <li>Design a grove 0.5 mm from the edge, and carefully cut out a slope going down to the grove. </li>
 <li>Cut out one or two full-thickness nicks into the tubing. These will help manipulating the cuff during graft preparation and transplantation.</li>
 <li>Cut the cuff at 4 mm from its edge. </li>
</ol>

<ol>
	<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> J. Vis. Exp. (65), e4143 (2012).</li><li>Hori, T., 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=Comprehensive+and+innovative+techniques+for+liver+transplantation+in+rats:+A+surgical+guide.">Comprehensive and innovative techniques for liver transplantation in rats: A surgical guide.</a> World Journal of Gastroenterology., WJG. 16, 3120 (2010).</li><li>Kashfi, 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+review+of+various+techniques+of+orthotopic+liver+transplantation+in+the+rat.">A review of various techniques of orthotopic liver transplantation in the rat.</a> Transplantation Proceedings. 37 (1), 185-188 (2005).</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 (1979).</li><li>Oldani, G., 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+novel+technique+for+rat+liver+transplantation+using+Quick+Linker+system:+a+preliminary+result.">A novel technique for rat liver transplantation using Quick Linker system: a preliminary result.</a> Journal of Surgical Research. 149, 303-309 (2008).</li></ol>

The authors have nothing to disclose.

The present report describes the design and building of the quick-linker system for easier rat liver transplantation.
Rat liver transplantation is a popular model with 125 to 180 related papers published yearly since the early 1990's (<a href="http://www.pubmed.com" target="_blank">www.pubmed.com</a>). It allows for a wide range of experiments including the exploration of immunological and graft preservation issues. The use of the "quick-linker" kit allows for easier and quicker anastomoses with guaranteed intima-to-intima contact. A detailed description and discussion of the technical steps of a rat liver transplantation have been described previously 1. 
The quick-linker kit can be built at low price from common laboratory material. The main challenge encountered has been related the breakdown of one handle during the final brushing, and that problem has been solved by setting the narrowest part of the handle ring at 0.5 mm (Figures 1 and 2 show appropriate sizes). Regarding the blade quality, only stainless steel should be considered. Although slightly less flexible compared to the average carbon steel blade, stainless steel blades do not develop rust, allowing a lifetime use of the handles. 
Of note, the design of the current handles and cuffs is the result of a process, which has led to the proposed sizes, allowing for the most efficient anastomoses. Cuff diameters vary according to the rat weights (Table 1). Minimal variations can be tolerated in terms of length, with a usual length of about 4 mm (Figures 3 and 4). 
Regarding the approximator, the selected Kocher's forcep should have handles at least 4 cm from the fulcrum. This allows for a safe closure of the rack outside the animal's abdomen. 
Overall, the proposed device can allow easier vein anastomoses for the transplantation of liver and other types of grafts in rats.

A technique of orthotopic rat liver transplantation has been shown in a recent Journal of Visualized Experiments paper 1. Portal vein and infra-hepatic vena cava anastomoses were performed using the quick-linker assisted cuff technique. This technique was chosen among other revisions of the original Kamada's technique 2-4, as it allows for easier and quicker anastomoses, results in a better hemodynamic, and can be implemented with a shorter learning curve 1,5.
The quick-linker technique requires the use of a handle, an approximator and cuffs, all of which can be designed from common laboratory material according to the technique described in the present report.

<table><tbody><tr><td>Porta cuff</td><td>altecweb or smiths-medical</td><td>Coil 01-96-1729&lt;br/&gt; 800/100/420</td><td>Bore : variable &lt;br/&gt; Wall : 0.30 mm &lt;br/&gt; Length: 3.8-4.2 mm&lt;br/&gt; &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></tr><tr><td>IVC cuff</td><td>altecweb or smiths-medical</td><td>Coil 01-96-1733&lt;br/&gt; 800/100/540</td><td>Bore : variable &lt;br/&gt; Wall : 0.30 mm&lt;br/&gt; Length: 3.8-4.2 mm&lt;br/&gt; &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></tr><tr><td>Mini-drill </td><td>Dremel</td><td>100 series</td><td>35,000 rpm&lt;br/&gt; &lt;a href=&quot;http://dremel.com&quot; target=&quot;_blank&quot;&gt;dremel.com&lt;/a&gt;</td></tr><tr><td>Diamond wheel point</td><td>Dremel</td><td>7103</td><td>2 mm&lt;br/&gt; &lt;a href=&quot;http://dremel.com&quot; target=&quot;_blank&quot;&gt;dremel.com&lt;/a&gt;</td></tr><tr><td>Diamond wheel point</td><td>Dremel</td><td>7105</td><td>4.4 mm&lt;br/&gt; &lt;a href=&quot;http://dremel.com&quot; target=&quot;_blank&quot;&gt;dremel.com&lt;/a&gt;</td></tr><tr><td>Diamond wheel point</td><td>Dremel</td><td>7144</td><td>2.4 mm&lt;br/&gt; &lt;a href=&quot;http://dremel.com&quot; target=&quot;_blank&quot;&gt;dremel.com&lt;/a&gt;</td></tr><tr><td>Cut-off wheels</td><td>Dremel</td><td>420</td><td>dia: 23.8 mm&lt;br/&gt; thickness: 0.5 mm&lt;br/&gt; &lt;a href=&quot;http://dremel.com&quot; target=&quot;_blank&quot;&gt;dremel.com&lt;/a&gt;</td></tr><tr><td>Quick-linker handles</td><td>dltchiropody (DLT)</td><td>PAR 10</td><td>modified scalpel blades&lt;br/&gt; &lt;a href=&quot;http://dltchiropody.co.uk&quot; target=&quot;_blank&quot;&gt;dltchriopdy&lt;/a&gt;</td></tr><tr><td>Quick-linker approximator</td><td>pharmap</td><td>UD-04-275</td><td>modified Kocher&#039;s forceps&lt;br/&gt; &lt;a href=&quot;http://pharmap.ch&quot; target=&quot;_blank&quot;&gt;pharmap.ch&lt;/a&gt;</td></tr></tbody></table>

manufacturing devices and instruments for easier rat liver transplantation

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This technique allows for easier venous cuff-anatomoses after a reasonable learning curve. The device is composed of two handles, which are carved out from scalpel blades, one approximator, which is obtained by modifying Kocher's forceps, and cuffs designed from fine-bore polyethylene tubing. The whole process can be performed at a low-cost using common laboratory material. The present report provides a step-by-step protocol for the design of the required pieces and includes stencils.

Five kits, including five approximators and 11 handles have been manufactured. One handle broke during the final brushing. This problem was due to the fact that the ring of the handle was too narrow, and it has been solved by setting the narrowest part of the handle ring at 0.5 mm. No problems occurred during the shaping of the approximators.
<table border="1">
<tbody>
 <tr>
 <td>Rat weight 
 (g)</td>
 <td>Porta handle (inner dia) 
 (mm)</td>
 <td>Porta cuff 
 (outer dia) 
 (mm)</td>
 <td>IVC handle (inner dia) 
 (mm)</td>
 <td>IVC cuff 
 (outer dia) 
 (mm)</td>
 </tr>
 <tr>
 <td>120-180</td>
 <td>2.2</td>
 <td>1.7</td>
 <td>2.6</td>
 <td>2.08</td>
 </tr>
 <tr>
 <td>180-270</td>
 <td>2.6</td>
 <td>2.08</td>
 <td>3.4-3.7</td>
 <td>3.0-3.25</td>
 </tr>
 <tr>
 <td>&gt;270</td>
 <td>2.9</td>
 <td>2.42</td>
 <td>4.2</td>
 <td>3.7</td>
 </tr>
 </tbody>
</table>
Table 1.
<img src="/files/ftp_upload/50380/50380fig1.jpg" alt="Figure 1" fo:content-width="6in" fo:src="/files/ftp_upload/50380/50380fig1highres.jpg"/> 
Figure 1. Porta handle stencil. If printed with a scale 1:1 can be used to guide the handle shaping. <a href="https://www.jove.com/files/ftp_upload/50380/50380fig1large.jpg" target="_blank">Click here to view larger figure</a>.
<img src="/files/ftp_upload/50380/50380fig2.jpg" alt="Figure 2" fo:content-width="6in" fo:src="/files/ftp_upload/50380/50380fig2highres.jpg"/> 
Figure 2. IVC handle stencil. If printed with a scale 1:1 can be used to guide the handle shaping. <a href="https://www.jove.com/files/ftp_upload/50380/50380fig2large.jpg" target="_blank">Click here to view larger figure</a>.
<img src="/files/ftp_upload/50380/50380fig3.jpg" alt="Figure 3" fo:content-width="4.6in" fo:src="/files/ftp_upload/50380/50380fig3highres.jpg" /> 
Figure 3. Porta and IVC cuffs lateral view. Graft's v.porta should be secured to the central groove, while recipient's vessel to the lateral one. This expedient allows for optimal intima-to-intima contact.
<img src="/files/ftp_upload/50380/50380fig4.jpg" alt="Figure 4" fo:content-width="4.1in" fo:src="/files/ftp_upload/50380/50380fig4highres.jpg"/> 
Figure 4. Porta and IVC cuffs front view. Calipers suitable for rats weighing 180-270 g are represented.

Watch this Scientific Journal Video about Manufacturing Devices and Instruments for Easier Rat Liver Transplantation at JoVE.com

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

We describe the design of the &#x201C;quick-linker&#x201D; device for easier orthotopic rat liver transplantation.

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This ...

manufacturing-devices-instruments-for-easier-rat-liver

Research

JoVE Journal

Bioengineering

8.7K Views.  University of Geneva Hospitals. We describe the design of the “quick-linker” device for easier orthotopic rat liver transplantation.

Video: Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

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 ...

Medicine

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

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

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.

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device

Decellularization and recellularization of parenchymal organs may enable the generation of functional organs in vitro, and several protocols for rodent liver decellularization have already been published. We aimed to improve the decellularization process by construction of a proprietary perfusion device enabling selective perfusion via the portal vein and/or the hepatic artery. Furthermore, we sought to perform perfusion under oscillating surrounding pressure conditions to improve the homogeneity of decellularization. The homogeneity of perfusion decellularization has been an underestimated factor to date. During decellularization, areas within the organ that are poorly perfused may still contain cells, whereas the extracellular matrix (ECM) in well-perfused areas may already be affected by alkaline detergents. Oscillating pressure changes can mimic the intraabdominal pressure changes that occur during respiration to optimize microperfusion inside the liver. In the study presented here, decellularized rat liver matrices were analyzed by histological staining, DNA content analysis and corrosion casting. Perfusion via the hepatic artery showed more homogenous results than portal venous perfusion did. The application of oscillating pressure conditions improved the effectiveness of perfusion decellularization. Livers perfused via the hepatic artery and under oscillating pressure conditions showed the best results. The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion set-ups to improve decellularization and recellularization experiments in rat livers.

The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion set-ups to improve decellularization and recellularization experiments in rat livers.

Decellularization and recellularization of parenchymal organs may enable the generation of functional organs in vitro, and several protocols for rodent ...

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

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the failing native liver has regenerated.1-3 The APOLT-method, the orthotopic implantation of auxiliary segments- averts most of the technical problems. However this method necessitates extensive resections of both the native liver and the graft.4 In 1998, Erhard developed the heterotopic auxiliary liver transplantation (HALT) utilizing portal vein arterialization (PVA) (Figure 1). This technique showed promising initial clinical results.5-6 We developed a HALT-technique with flow-regulated PVA in the rat to examine the influence of flow-regulated PVA on graft morphology and function (Figure 2).
A liver graft reduced to 30 % of its original size, was heterotopically implanted in the right renal region of the recipient after explantation of the right kidney.&#160; The infra-hepatic caval vein of the graft was anastomosed with the infrahepatic caval vein of the recipient. The arterialization of the donor&#8217;s portal vein was carried out via the recipient&#8217;s right renal artery with the stent technique. The blood-flow regulation of the arterialized portal vein was achieved with the use of a stent with an internal diameter of 0.3 mm. The celiac trunk of the graft was end-to-side anastomosed with the recipient&#8217;s aorta and the bile duct was implanted into the duodenum. A subtotal resection of the native liver was performed to induce acute hepatic failure. 7
In this manner 112 transplantations were performed. The perioperative survival rate was 90% and the 6-week survival rate was 80%. Six weeks after operation, the native liver regenerated, showing an increase in weight from 2.3&#177;0.8 g to 9.8&#177;1 g. At this time, the graft&#8217;s weight decreased from 3.3&#177;0.8 g to 2.3&#177;0.8 g.
We were able to obtain promising long-term results in terms of graft morphology and function. HALT with flow-regulated PVA reliably bridges acute hepatic failure until the native liver regenerates.

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.

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

Rat Heterotopic Abdominal Heart/Single-lung Transplantation in a Volume-loaded Configuration

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft involves anastomosis of donor inferior vena cava (IVC) to recipient IVC, and donor ascending aorta (Ao) to recipient abdominal Ao. The right upper and middle lung lobes are preserved and function as conduits for blood flow from right heart to left heart.
There are several advantages to using this technique, and it lends itself to a broad range of applications. Because the graft is transplanted in a configuration that allows for dyamic volume-loading, cardiac function may be directly assessed in vivo. The use of pressure-volume conductance catheters permits characterization of load-dependent and load-independent hemodynamic parameters. The graft may be converted to a loaded configuration by applying a clamp to the recipient&#8217;s infra-hepatic IVC. We describe modified surgical techniques for both donor and recipient operations, and an ideal myocardial protection strategy. Depending on the experimental aim, this model may be adapted for use in both acute and chronic studies of graft function, immunologic status, and variable ventricular loading conditions. The conducting airways to the transplanted lung are preserved, and allow for acute lung re-ventilation. This facilitates analysis of the effects of the mixed venous and arterial blood providing coronary perfusion to the graft.
A limitation of this model is its technical complexity. There is a significant learning curve for new operators, who should ideally be mentored in the technique. A surgical training background is advantageous for those wishing to apply this model. Despite its complexity, we aim to present the model in a clear and easily applicable format. Because of the physiologic similarity of this model to orthotopic transplantation, and its broad range of study applications, the effort invested in learning the technique is likely to be worthwhile.

We describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The transplant configuration results in a partially loaded graft circulation, allowing direct functional assessment. This model may be employed for acute or chronic studies of function and immunologic status of the transplanted graft.

Herein, we describe a novel technique for heterotopic abdominal heart-lung transplantation (HAHLT) in rats. The configuration of the transplant graft ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

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

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

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

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.

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

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.

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 ...

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

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

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&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Liver Transplantation in Rats and Analyzing Allograft Patency

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

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat liver transplantation (LTx) protocols typically use an orthotopic model where the recipients&#39; native liver is removed and replaced with a donor liver. This technically demanding surgical procedure requires advanced micro-surgical skills and is further complicated by lengthy anhepatic and lower body ischemia times. This prompted the development of a less complicated heterotopic method that can be performed faster with no anhepatic or lower body ischemia time, reducing post-surgery stress for the recipient animal.
This heterotopic LTx protocol includes two main steps: excising the liver from the donor rat and transplanting the whole liver into the recipient rat. During the excision of the donor liver, the surgeon ligates the supra-hepatic vena cava (SHVC) and hepatic artery (HA). On the recipient side, the surgeon removes the left kidney and positions the donor liver with the portal vein (PV), infra-hepatic vena cava (IHVC), and bile duct facing the renal vessels. Further, the surgeon anastomoses the recipient&#39;s renal vein end to end with the IHVC of the liver and arterializes the PV with the renal artery using a stent. A hepaticoureterostomy is utilized for biliary drainage by anastomosing the bile duct to the recipient&#39;s ureter, permitting the discharge of bile via the bladder.
The average duration of the transplantation was 130 min, cold ischemia duration was around 35 min, and warm ischemia duration was less than 25 min. Hematoxylin and eosin histology of the auxiliary liver from syngeneic transplants showed normal hepatocyte structure with no significant parenchymal alterations 30 days post-transplant. In contrast, 8-day post-transplant allogeneic graft specimens demonstrated extensive lymphocytic infiltration with a Banff Schema rejection activity index score of 9. Therefore, this LTx method facilitates a low morbidity rejection model alternative to orthotopic LTx.

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.

Small animal transplant models are indispensable for organ tolerance studies investigating feasible therapeutic interventions in preclinical studies. Rat ...

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