A Microsurgical Modelo Novel para heterotópico, em bloco da parede torácica, Thymus, e Transplante Cardíaco em Ratos
Summary
To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.
Abstract
Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.
Introduction
Cardiac transplantation is the treatment of choice for end-stage heart failure. Both technical advancements and pharmacological innovations have propelled the field to early graft acceptance rates above 90%.12,13 Despite this, 60-80% 5-year graft survival is at a standstill and chronic rejection, characterized by transplant vasculopathy, remains inevitable.14-16 Furthermore, patients are subjected to multiple surgical procedures and lifelong immunosuppression, which are associated with chest wall deformities and medical sequelae and toxicities, respectively. The need for innovative approaches to extend allograft survival, minimize the immunosuppressive requirements, and offer reconstructive options for anatomical deformities is pressing.
Vascularized composite allotransplantation offers a unique strategy for improving heart transplant outcomes both from an immunological aspect as well as a reconstructive perspective.17 Vascularized composite allografts are also unique in a way that they have an inherent source of donor-derived hematopoietic stem cells which has shown a favorable ability to reduce immunosuppression and induce and sustain mixed chimerism.1-8 Additionally, co-transplantation of the thymus has shown to prolong survival of both, solid organ transplants and vascularized composite allografts.2,9-11 Combining these strategies with heart transplantation offers a novel solution to the aforementioned challenges facing heart transplantation.18
Murine models serve as excellent platforms for mechanistic in vivo investigation because of the availability of antibodies and well-defined inbred and knockout strains.19-21 Although heart transplantation in mice is commonly studied using a heterotopic intraabdominal microsurgical suture transplant model22-25, a heterotopic, cervical, non-suture cuff technique model has shown to be extremely replicable, reliable, and carries fewer rates of thrombosis.19,26,27 The goal of this study is to develop a heterotopic en bloc osteomyocutaneous chest wall, thymus, and heart transplant technique in mice to study the immunological mechanisms of combined solid organ and vascularized composite allotransplantation using a cervical non-suture cuff technique. This cluster allograft is perfused through the anastomosis of the donor descending aorta to the right common carotid artery and the donor pulmonary artery to the right external jugular vein. Preservation of the internal thoracic vessels and associated thymus branches is paramount to perfusing the chest wall (sternum, ribs, muscles, and skin) and thymus.
Protocol
Representative Results
Discussion
Há uma infinidade de fenómenos que factor para a investigação imunológica de alotransplante, que incluem mas não estão limitados a mecanismos de rejeição aguda e crónica, apresentação directa e indirecta antigénio, sensibilização destinatário, ou a indução de quimerismo misto. 19 Modelos animais se tornaram o padrão-ouro para o estudo da imunologia do transplante, e mouse modelos são popularmente implementada devido ao seu baixo custo, disponibilidade de camundongos transgênicos e knockou…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work was funded by the American Association of Plastic Surgeons 2014 Academic Scholar Award.
Materials
| Euro-Collins Solution | The solution is not commercially purchased but rather prepared in the laboratory. To make a 500ml solution add the ingredient listed below to a 330ml of double distilled water. Mix well, and then fill in the rest of the 170ml of double distilled water into the solution to a final volume of 500ml. Ingredients: 1.02g KH2PO4, 3.66g K2HPO4, 0.56g KCl, 0.42g NaHCO3, and 17.52g of glucose. |
||
| Suture | Ethilon | MWI 72667 | 6-0 Ethilon |
| Suture | Microsurgery Instruments, Inc. | 10-0 Nylon | 10-0 Nylon |
| Polyimide Cuff Vein (21G) | Vention Medical | 141-0043 | http://www.ventionmedical.com/products-and-services/polyimide-tubing/ |
| Polyimide Cuff Artery (24G) | Vention Medical | 141-0027 | http://www.ventionmedical.com/products-and-services/polyimide-tubing/ |
| Soft plastic tip catheter | Terumo | SR*OX2419CA | 24G x 3/4" |
| Microsurgical dilator | S&T | D-5a.1 | Dilator, 11cm, FH, 0.1mm AT10d |
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