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

Abstract

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 wellstandardized 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. Video LinkThe video component of this article can be found at https://www.jove.com/video/4376/ Protocol Basic Techniques and Common Procedures1. All procedures are carried out under clean but nonsterile conditions. 2. Once the abdomen of the rat is opened, all procedures are performed under a surgical microscope at a magnification of 16x. Exceptions are the ex vivo liver resection, which is performed at 10x, and the following procedures, which are performed at 25x: insertion of a stent into the bile duct and the hepatic artery, and the reconstruction of the hepatic artery, the infrahepatic vena cava (IHVC), and the bile duct. 3. Cotton swabs are used for gentle manipulation of organs, blunt dissection of tissues, and compression hemostasis. Gauze swabs (5 x 5 cm) soaked with lactated Ringer solution are used to retract the liver or intestines, and to keep organs moist. The Satinsky clamp can be used for retraction of gauze-covered intestines toward the left or the tail of the ...

Improving Research Practice in Rat Orthotopic and Partial Orthotopic Liver Transplantation: A Review, Recommendation, and Publication Guide

“…Table 1 shows the numerous creative technical solutions elaborated to optimize graft rearterialization in rat OLT and P-OLT [8,12,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39]. …”

“…Besides these well-designed multistep training programs, further reports are available in the literature that provide a detailed, pragmatic surgical guide for researchers who are aiming to master this model [14,20,37,47]. …”

“…Following this pretraining, the observation of skilled microsurgeons and repeatedly watching training videos - prepared by our working group - represent the next stage. One of these didactic and detailed training videos was recently published by Nagai et al [37] in the Journal of Visualized Experiments .…”

Defining Standards in Experimental Microsurgical Training: Recommendations of the European Society for Surgical Research (ESSR) and the International Society for Experimental Microsurgery (ISEM)

“…They showed a high satisfaction, especially among students residing far from their training centers. Today, free-of-charge video platforms are widely available and even scientific video journals are gaining ground; therefore, the role of quality and didactic training videos in surgical training and in its international standardization is highly emphasized [23, 26, 27]. …”

Operative Techniques, Treatment, and Precaution of Common Complications of Orthotopic Rat Liver Transplantation

“…Third, postoperative 24 h death of rats was primarily caused by blood leakage and errhysis of anastomotic stoma of SHVC. As reported [3], postoperative survival rate is affected by three key factors: amount of blood loss, operation time, and adequacy of reconstruction of each vessel. The key point for decreasing this complication includes using tunica intima-to-tunica intima continuous suture, keeping proper intervals between needles, maintaining proper tension on suture line, and mastering consummate techniques of saturation.…”