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In This Article

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

Summary

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

Abstract

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.

Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Introduction

Over the past six decades, solid organ transplantation has evolved from an experimental procedure to a standard of care for the treatment of end-stage organ failure1. Due to the improvement of antimicrobial agents, surgical techniques and advancement in immunosuppressive regiments, the early success rate of solid organ transplantation have significantly increased over the past decades2.

However, long-term graft survival rates have not significantly improved in the same manner3. The development of CAV is the major factor limiting long-term survival4

Protocol

All experiments were performed according to the guidelines of the German animal welfare act (TierSchG.) (AZ: 55.2-1-54-2532.Vet_02-80-2015).

1. Animal housing

  1. For experiments, use male C57BL/6 and BALB/c mice weighing 20-25 g with C57BL/6 mice as the recipient animals and BALB/c mice as the donor animals.
  2. Purchase the animals and house in a barrier pathogen-free facility, in accordance with the FELASA guidelines for health monitoring12.
  3. K.......

Representative Results

In the fully MHC-mismatch transplantation model, a concentric neointimal layer can be seen 4 weeks after transplantation (Figure 2). This layer consists primarily of vascular smooth muscle cells as immunohistological staining for SM22 (a selective marker for mature vascular smooth muscle cells) revealed. As stated before, these vascular smooth muscle cells are pathognomonic for lesions seen in chronic allograft vasculopathy. For further analyses, aortic segments should be sectioned and stain.......

Discussion

Chronic allograft vasculopathy is the major cause of late graft loss after solid organ transplantation of the heart and likely renal and lung allografts8. Thus far, no causal therapeutic regimen could be developed in order to prevent the formation of CAV.

The pathophysiology of CAV is multifactorial and involves immunological and non-immunological aspects16. The use of rodent models in transplantation have been essential in understanding the unde.......

Acknowledgements

None.

....

Materials

NameCompanyCatalog NumberComments
Balb-c Mice (H2-d)Charles RiverStrain# 028Donor animal
Bipolar cautery systemERBEICC 50 / 20195-023Bipolar cautery
C57BL/6J (H-2b)Charles RiverStrain# 027Recipient animal
Halsey Needle HoldersFST12501-12Needle Holder
Halsted-Mosquito ForcepsAESCULAPBH111RCurved Clamp
Medical Polyimide TubingNordson MEDICAL141-0031Cuff-Material
Micro SerrefinesFST18055-04Micro Vessel Clip
Micro-Adson Forceps (serrated)FST11018-12Standard Forceps
Micro-Serrefine Clamp Applying ForcepsFST18057-14Clipapplicator
S&T Forceps - SuperGrip Tips (Angled 45°)S&T00649-11Fine Forceps
S&T Vessel Dilating Forceps - Angled 10° (Tip diameter 0.2 mm)S&T00125-11Vesseldilatator
Schott VisiLED SetSchottMC 1500 / S80-55Light
Stereoscopic microscopeZEISSSteREO Discovery.V8Microscope
Student Fine Scissors / Surgical Scissors - Sharp-BluntFST91460-11 / 14001-12Standard Sissors
Vannas-Tübingen Spring Scissors (curved, 8.5 cm)FST15004-08Microsissors (curved)
Vannas-Tübingen Spring Scissors (straight, 8.5 cm)FST15003-08Microsissors (straight)

References

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MurineCervical Aortic TransplantationNon suture Cuff TechniqueHeterotopic Aortic TransplantationCommon Carotid ArteryVascular SurgeryMicrosurgical Technique

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