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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

Abstract

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Introduction

Abbott et al. published1 the first description of heterotopic abdominal heart transplantation in rats in 1964. These surgical techniques were refined and simplified by Ono et al. in 19692. Corry et al. first described a method for heterotopic abdominal heart transplantation in mice in 1973; similar to the previously reported rat models, this involved engraftment into the host's abdomen with revascularization by end-to-side anastomoses of the donor's pulmonary artery and ascending aorta to the recipient's inferior vena cava and abdominal aorta, respectively3. Heterotopic cervical heart ....

Protocol

All animal handling procedures were conducted in compliance with the NIH Care and Use of Laboratory Animals guidelines and approved by the Animal Studies Committee at Washington University School of Medicine. Hearts from C57BL/6 (B6) and BALB/c mice (weighing 20-25 g) were transplanted into gender-matched B6 recipients (6-8 weeks of age). The mice were obtained from commercial sources (see Table of Materials). Syngeneic transplants were performed to evaluate cellular responses related to ischemia-reperfu.......

Representative Results

This mouse cervical heterotopic cardiac transplantation model has been utilized to perform over 1,000 transplants in our laboratory, with a survival rate of approximately 97%. The success rate is slightly higher than previous reports using other cervical heterotopic heart transplantation techniques in mice10,11,20. This could potentially be attributed to the larger 20 G cuff placed on the donor pulmonary artery to ensure ample r.......

Discussion

Utilizing this technique, mouse heterotopic cervical cardiac transplantation can be performed in less than 40 min by an experienced microsurgeon and in approximately 60 min by an entry-level microsurgeon. While cervical heart transplantation has been studied in numerous animal models, a mouse model remains the gold standard due to multiple well-defined genetic strains, genetic alteration capabilities, and the availability of numerous reagents, including monoclonal antibodies24. The technique descr.......

Acknowledgements

DK is supported by National Institutes of Health grants 1P01AI116501, R01HL094601, R01HL151078, Veterans Administration Merit Review grant 1I01BX002730, and The Foundation for Barnes-Jewish Hospital.

....

Materials

NameCompanyCatalog NumberComments
6-0 braided silk tiesHenry Schein Inc7718729
0.75% Providone iosine scrubPriority Care IncNDC 57319-327-0
10-0 nylon sutureSurgical Specialties CorporationAK-0106
655-nm nontargeted Q-dotsInvitrogenQ21021MP
70% EthanolPharmco Products Inc111000140
8-0 braided silk tiesHenry Schein Inc1005597
Adson forcepsFine Science Tools Inc91127-12
BALB/c and C57BL/6 mice (6-8 weeks)Jackson Laboratories
Bipolar coagulatorValleylab IncSurgII-20, E6008/E6008B
Carprofen (Rimadyl) injectionTranspharm35844
Carprofen (Rimadyl) oral chewable tabletTranspharm38995/37919
Custom-built 2P microscope running ImageWarp acquisition softwareA&B Software
Dumont no. 5 forcepsFine Science Tools Inc11251-20
Fine vannas style spring scissorsFine Science Tools Inc15000-03
GraphPad Prism 5.0Sun Microsystems Inc.
Halsey needle holderFine Science Tools Inc91201-13
Halsted-Mosquito clamp curved tipFine Science Tools Inc91309-12
Harvard Apparatus mouse ventilator model 687Harvard ApparatusMA1 55-0001
Heparin solution (100 U/mL)Abraxis Pharmaceutical Products504031
ImarisBitplane
Ketamine (50 mg/kg)Wyeth206205-01
Microscope—Leica Wild M651 × 6–40 magnificationLeica Microsystems
Moria extra fine spring scissorsFine Science Tools Inc15396-00
Ohio isoflurane vaporizerParkland ScientificV3000i
QdotsThermoFisher1604036
S&T SuperGrip Forceps angled tipFine Science Tools Inc00649-11
S&T SuperGrip Forceps straight tipFine Science Tools Inc00632-11
Sterile normal saline (0.9% (wt/vol) sodium chlorideHospira IncNDC 0409-4888-20
Sterile Q-tips (tapered mini cotton tipped 3-inch applicators)Puritan Medical Company LLC823-WC
Surflow 20 gauge 1/4-inch Teflon angiocatheterTerumo Medical CorporationSR-OX2032CA
Surflow 24 gauge 3/4-inch Teflon angiocatheterTerumo Medical CorporationR-OX2419CA
ThermoCare Small Animal ICU System (recovery settings 3 L/min O2, 80 °C, 40% humidity)Thermocare Inc
VetBondSanta Cruz Biotechnology SC361931NC0846393
Xylazine (10 mg/kg)Lloyd Laboratories139-236

References

  1. Abbott, C. P., Lindsey, E. S., Creech, O., Dewitt, C. W. A technique for heart transplantation in the rat. The Archives of Surgery. 89, 645-652 (1964).
  2. Ono, K., Lindsey, E. S. Improved technique of heart transplantation in rats.

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