A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Summary

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

Abstract

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients' long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end anastomosis to the recipient's aorta and inferior vena cava using stitches. In this model, the donor's heart is implanted by end-to-end anastomosis to the recipient's carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

Introduction

Heart transplantation has become the standard treatment for terminal heart failure. More than 5,500 heart transplantations per year are performed in the organizations registered under the International Society for Heart and Lung Transplantation. Among the allogeneic heart transplant recipients, the 1-year rejection rate is still >10%, while the 3-year rejection rate increased to 36%^1,2. However, effective prophylactic treatments for patients with cardiac allograft rejection are lacking. Therefore, animal model studies are warranted that elucidate the physiological mechanisms underlying the immunological an....

Protocol

All Animal experiments were performed in compliance with the Guide for the Care and Use of Laboratory Animals, Eight Edition, National Research Council (US) 2011. Procedures involving animal subjects have been approved by the Animal Care and Use Committee of the Chongqing University Cancer Hospital, Chongqing, China. Male BALB/c and C57BL/6 mice weighing 20-30 g, obtained from commercial sources (see Table of Materials), were used for allogeneic heart transplantation study. The C57BL/6 mice were use.......

Discussion

The mouse heart transplantation model contributes to the investigation of rejection mechanisms after heart transplantation, contributing to the development of unique approaches to improve the long-term survival of cardiac allograft recipients. However, heart transplantation in mice is a complex and challenging task, requiring a high level of microsurgery techniques, especially in vascular anastomosis^11,12,13. The mouse abdominal.......

Materials

Name	Company	Catalog Number	Comments
5-0 Polyamide Monofilament suture	B.Braun Medical Inc.	C3090954
8-0 Polyamide Monofilament suture	B.Braun Medical Inc.	C2090880
10-0 Polyamide Monofilament suture	B.Braun Medical Inc.	G0090781
22 G polyurethane cuff	B.Braun Medical Inc.	4251628-02
26 G polyurethane cuff	Suzhou Linhua Medical Instrument Co., LTD	REF383713
Anesthesia induction chamber	RWD Life Science Co., LTD	V100
Atraumatic microvascular clamp	Beyotime	FS500
BALB/c and C57BL/6 mice (20–30 g)	Centre of Experimental Animals (Army Medical University, Chongqing, China)
Buprenorphine	US Biological life Sciences	352004
Electrocoagulator	Guangzhou Runman Medical Instrument Co., LTD	ZJ1099
Gauze	Henan piaoan group Co., LTD	10210402
Heating pad	Guangzhou Dewei Biological Technology Co., LTD	DK0032
Heparin	North China Pharmaceutical Co., LTD	2101131-2
HTK solution	Shenzhen Changyi Pharmaceutical Co., LTD	YZB/Min8263-2013
Injection syringe (10 mL)	Shandong weigao group medical polymer Co., LTD	20211001
Isoflurane	RWD Life Science Co., LTD	21070201
Physiological saline	Southwest pharmaceutical Co., LTD	H50021610
Scalp needle	Hongyu Medical Group	20183150210
Shaver	Beyotime	FS600
Small animal anesthesia machine	RWD Life Science Co., LTD	R500
Surgical operation microscope	Tiannuoxiang Scientific Instrument Co. , Ltd, Beijing, China	SZX-6745
Swab	Yubei Medical Materials Co., LTD	21080274