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This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.
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.
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....
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.......
PV reconstruction is successful when, upon unclamping the portal vein, there is no tortuosity, and the liver is uniformly perfused. Anhepatic time should be under 20 min, as anhepatic times exceeding 25 min increase the risk of mouse mortality. IHIVC reconstruction is deemed successful if there is no blood regurgitation from the graft.
Storing the graft at cold temperatures for 1 h using organ preservation solution results in a serum alanine aminotransferase level of approximately 2,000 U/L at.......
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 connection2,5, some minor p.......
This work was supported by 2022 JST basic research (The Japanese Society for Transplantation).
....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 |
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