Improved Cuff Technique for Establishing a Mouse-Rat Heterotopic Cardiac Xenotransplantation Model

Abstract

Background: The small animal model of cardiac transplantation is the most common model in organ transplantation studies. The cervical heterotopic transplantation is widely performed because this allows for direct observation of the graft heartbeat and contributes to early prediction of graft rejection.

Objective: A mouse-rat cervical heterotopic cardiac xenotransplantation model was modified with respect to the anesthesia method, cardiac graft harvesting method, and perioperative treatment. These improvements ensure the stability and reliability of xenotransplantation models for in vivo studies of immune-mediated graft rejection.

Methods: After establishing isoflurane inhalation anesthesia, the donors' hearts were harvested. The experimental method involved separate ligation of the left and right superior venae cavae; the other blood vessels were ligated in a cluster. Both the donor and recipient animals were placed on a heating pad intraoperatively to maintain a body temperature of 37-40 °C. The model establishment was divided into 3 stages: practice, stabilization, and stereotyping. The surgical success rate and operation time were recorded. Specimens were harvested at different time points for histopathological examination.

Results: The anesthetic effect of isoflurane was well maintained, and no animals died of adverse anesthetic events. Body temperature was maintained at 37-40 °C which effectively shortened the time to restoration. The modification of the cardiac graft harvesting method is conducive to rebeating of the donor heart. The success rates in the stabilization and stereotyping stages were significantly higher than that in the practice stage (P < .05). The operation time in the stabilization and stereotyping stages were significantly shorter than those in the practice stage (P < .05). Histopathological examination revealed thrombosis formation, interstitial hemorrhage, and inflammatory cell infiltration in the donor hearts.

Conclusion: Our findings suggest that the mouse-rat cervical heterotopic cardiac xenotransplantation model is the ideal animal model for studying xenograft rejection.