Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

Summary

This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.

Abstract

Mouse orthotopic liver transplantation is an effective methodology for investigating the underlying mechanisms of liver ischemia and reperfusion injury. However, the technical challenges pose a barrier to utilizing this valuable experimental model and passing on these skills to the next generation. The most challenging aspect of this procedure is vascular reconstruction, including the portal vein (PV), infrahepatic inferior vena cava (IHIVC), and suprahepatic inferior vena cava. The use of plastic cuffs, rather than sutures, allows for smoother PV and IHIVC reconstruction. Vessels are reconstructed by attaching a cuff made from an intravenous catheter to the tip of the graft vessel and interposing the cuff into the recipient vessel. The two most crucial aspects are properly visualizing the inner lumen of the vessel and avoiding the use of excessive force. Our aim is to provide a technical overview of vascular reconstructions using the cuff technique in recipient surgery. These technical tips for the cuff technique are expected to help microsurgeons facilitate vascular reconstruction and advance their research.

Introduction

Mouse orthotopic liver transplantation (MOLT) is an effective experimental method first reported in 1991¹. This experimental model, which utilizes genetically modified mice and various research reagents, has played a pivotal role in investigating warm and cold ischemia and reperfusion injuries. However, the model's high technical complexity has hindered the development of basic medicine for liver transplantation². MOLT involves three major steps: (1) retrieval of the liver from the donor mouse, (2) back table surgery, and (3) implantation of the liver into the recipient. Among these recipient procedures, vascular ana....

Protocol

The experimental protocol was approved by the Animal Experimentation Committee at Kyoto University. The study utilized C57BL/6 mice, aged over 10 weeks and weighing between 25 g and 30 g, obtained from a commercial source (see Table of Materials). All animals were anesthetized with 2.5% isoflurane (following institutionally approved protocols), maintained under specific pathogen-free conditions, and all experimental procedures were conducted in compliance with Kyoto University's Regulations on Animal.......

Discussion

Learning vascular reconstruction is the most challenging aspect of achieving successful MOLT. The cuff's quality significantly influences the reconstruction's difficulty, given the small size of mice⁵. This article provides a detailed protocol for cuff preparation, attachment, and reconstruction.

While there are no major differences from previous reports regarding cuff preparation and connection^2,5, some minor p.......

Materials

Name	Company	Catalog Number	Comments
16 G intravenous catheter	TERUMO	SR-FF2032	IHIVC cuff
20 G intravenous catheter	TERUMO	SR-FF1651	PV cuff
8-0 braid silk	Natsume Seisakusho	CR9-80B2	8-0 silk
Belzer UW Cold Storage Solution	Astellas		Organ preservation fluid
Bulldog clamp	B BRAUN	FB329R	Bulldog clamp
C57BL/6 mice	Oriental Bio Service
Isoflurane inhalation solution	Viatris		Anesthesic
Micro Blunted Tips 0.1 mm x 0.06 mm	F.S.T	11253-20	Straight microforceps
Micro Serrefine Clamp Applicator with Lock	F.S.T	18056-14	Vessel clip applicator
Micro Serrefines	F.S.T	18055-4	Vessel clip
No.11 Spare Blades	FEATHER Safety Razor	11	Blades
Ophthalmic scissor, round handle	B BRAUN	FD103R	Microscissor
Plastic rectangular-shaped container	Daiso		10 cm long, 15 cm wide and 6 cm high
SuperGrip Tips	F.S.T	00649-11	Curved microforceps
SZX7	Olympus	SZX7	Microscope