Details determining the success in establishing a mouse orthotopic liver transplantation model

Abstract

Liver transplantation (LT) is currently the only effective treatment option for end-stage liver disease. The importance of animal models in transplantation is widely recognized among researchers. Because of the well-characterized mouse genome and the greater diversity and availability of both genetically modified animals and research reagents, mouse orthotopic LT (MOLT) has become an ideal model for the investigation of liver biology, tissue injury, regulation of alloimmunity and tolerance induction, and the pathogenesis of specific liver diseases. However, due to its complicated and technically demanding procedure, the model has merely been used by only a few research groups in the world for years. For a new learner, training lasting at least a couple of months or even years is required. Most of the investigators have emphasized the importance of elaborate techniques and dedicated instruments in establishing a MOLT model, but some details are often neglected. The nontechnical details are also significant, especially for researchers who have little experience in mouse microsurgery. Here, we review and summarize the crucial technical and nontechnical details in establishing the model of MOLT based on scientific articles and our experience in six aspects: animal selection, anesthesia, perioperative management, organ procurement, back-table preparation, and implantation surgery. We aim to enable research groups to shorten the learning curve and implement the mouse LT procedure with high technical success.

Keywords: Anesthesia; Animal selection; Back-table preparation; Implantation surgery; Mouse orthotopic liver transplantation; Organ procurement; Perioperative management.

Figure 1

Necrosis of the hilar region due to ischemia of the proximal bile duct. A: Model diagram; B: General pathology; C: H&E staining.

Figure 2

A simplified method of cuff technique. A: Make sure the whole donor liver is immersed in water and rotate the donor liver floating in the dish so that the inferior surface faces upward; B: Pass the forceps through the lumen of the cuff to grasp the vessel so that the cuff is slipped over the vessel; C: Use forceps with a 45-degree angle to hold the extension of the cuff and use another curved round handle forceps to fold the end of the vessel over the cuff body to expose the inner endothelial surface and from one side to the other (orange arrow); D: Secure the PV(IHIVC) to the cuff by ligation. (CRITICAL STEP: Immerse donor liver under water so that the vessel wall naturally opens to facilitate vessel eversion).

Figure 3

Use a slipknot to temporarily occlude donor infrahepatic inferior vena cava. A: Demonstration of a slipknot: cross the fixed end (F) and the slip end (S), then keep F immobile and fold S to pass through the circle; B: Put the slipknot around the donor infrahepatic inferior vena cava (IHIVC) and fasten the slipknot. (Tip: After IHIVC anastomosis, pull S to loosen the slipknot).