Optimization of the Cuff Technique for Murine Heart Transplantation

Summary

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

Abstract

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

Introduction

The establishment of mouse heterotopic heart transplantation through end-to-end anastomosis within the abdomen in 1973 was a major milestone in basic transplant immunology research¹. This model provided an important and valid tool for analyzing the mechanisms of ischemia reperfusion injury², immunological rejection, and tolerance^3,4. However, the complex and time-consuming nature of the surgery as well as the potential for infections can result in severe perioperative abdominal adhesions and inflammatory reactions, resulting in a low efficiency for heterotopic he....

Protocol

Animals (BALB/c, C57BL/6, male, 8-12 weeks) are housed in a specific pathogen-free facility at the Xiamen University Laboratory Animal Center. C57BL/6 is used as recipient and BALB/c is used as donor. All the procedures are performed according to the Institutional Animal Care and Use Committee (IACUC) guidelines.

NOTE: A set of microsurgical instruments, including a micro scissor, micro straight forceps, micro curved forceps and micro needle holders, are necessary for the operation (Ta.......

Discussion

Mouse heart transplants models are important tools for transplant immunology research, as tools and materials for evaluating the immune mechanisms of this model and a large number of gene-modified mice are available. However, microsurgical technical challenges, such as vessels suture and eversion, have limited its widespread use. In the present study, we have investigated certain core technical challenges of murine heart transplantation and have obtained good outcomes. A critical step of the protocol the insertion of an .......

Materials

Name	Company	Catalog Number	Comments
Artery cuff	Self-made		Polyamide tube. diameter: 0.55 mm,length: 1.0 mm
Artery inner tube	Self-made		Polyamide tube. Diameter: 0.28mm
Micro curved forceps	Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory	WA3050	1/8 arc, 0.3-mm tip without a hook
Micro needle holders	Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory	WA2050	0.2-mm tip
Micro scissors	Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory	WA1050	Straight, blade length: 10 mm
Micro straight forceps	Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory	WA3060	0.15-mm tip without a hook
Scanlan Vascu-Statt Bulldog Clamps	Scanlan International Inc	1001-531	Clamping pressure 20–25 grams
Vein cuff	Self-made		Polyamide tube. diameter: 0.9 mm,length: 1.2 mm
Vein inner tube	Self-made		Polyamide tube. Diameter: 0.6 mm