原位后肢移植中的鼠标
Summary
这种新颖的模型在小鼠原位后肢移植，申请超微血管吻合的非缝合袖套法，提供了相关的血管化复合同种异体移植（VCA） 体内机械免疫学研究的有力工具。
Abstract
In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research.
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace “like with like” in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.
Introduction
The late nineties heralded the pioneering days of reconstructive transplantation with the first successful hand transplant performed in France in 1998. Since then, the use of VCAs for reconstruction of devastating tissue defects has been successfully employed in a wide spectrum of patients. To date, the world counts 76 recipients of 112 upper extremities as well as 31 faces 1-3. In addition, several other types of VCAs such as abdominal wall 4, larynx 5, trachea 6, vascularized joints 7, and even penis 8 have been performed. Furthermore, the live birth of a baby was recently reported after uterus transplantation 9. This growing world experience is indicative for how reconstructive transplantation has become a valid therapeutic option for patients suffering of significant functional tissue defects not amendable to conventional reconstructive and restorative surgery and treatment.
While the idea of replacing “like with like” sparked clinical enthusiasm, initial skepticism still prevails with regards to side effects of conventional high-dose immunosuppression required to maintain allografts and their function 10,11. However, as shown by seminal work of Lee et al., these composite grafts are less likely to reject than its individual components, and furthermore, some of the tissue components such as the vascularized bone compartment have fueled optimism as they might exert unique immunological effects onto the balance of immune acceptance and rejection 12.
Our group pioneered several microsurgical animal models for solid organ transplantation, as well as vascularized composite allotransplantation 13-19. Here we describe a novel surgical procedure using a non-suture cuff technique to perform super micro-vascular anastomosis in an orthotopic mouse hind limb transplantation model. This transplant model provides a useful tool for investigating immune acceptance and rejection mechanisms, as well as the role of individual tissue components, such as the vascularized bone marrow compartment, towards tolerance induction in the immunologically versatile setting of the mouse species. Additionally, the orthotopic placement of the limb opens the possibilities for nerve regeneration and functional outcome studies, which are critically important to the setting of VCA.
Protocol
Representative Results
Discussion
血管化复合异体移植，如上肢和面部移植毁灭性的组织缺损，已演变为对患者没有可修正传统的重建手术有效的治疗选择。在重建显微外科学领域的技术进步，以及与强效免疫和实体器官移植免疫调节疗法相当丰富的经验，现在可以在这个独特的患者人群3,21-长期移植存活。然而，对于移植的维护和生存需要长期免疫抑制显著的副作用仍然限制了这些提高生活而不是救命重建模式3,22,23…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项工作是由陆军，海军，美国国立卫生研究院，空军，VA和卫生事务的支持，支持AFIRM II的努力，在奖号W81XWH-13-2-0053。美国陆军医学研究收购活动，820钱德勒街，德特里克堡MD 21702-5014是颁发和管理采购办公室。意见，解释，结论和建议是那些作者的，不一定是由美国国防部批准。
笔者想在这项研究中感谢杰西卡IZZI，DVM，卡罗琳·加勒特，DVM和朱莉·沃森，DVM的出色兽医支持。
Materials
| Suture, 6-0 Nylon | MWI | 31849 | |
| Suture, 6-0 Polysorb | MWI | 72667 | |
| Suture, 10-0 Nylon | Aero Surgical | TK-107038 | |
| Polyimide Tubing, Size 25 | Vention Medical | 141-0023 | |
| Polyimide Tubing, Size 27 | Vention Medical | 141-0015 | |
| Microvascular Clamps (Single) | Synovis | 00396 | |
| Microvascular Clamps (Double) | Synovis | 00414 | |
| Micro-Scissors | Synovis | SAS-18 | |
| Micro-Forceps | Synovis | FRS-15 RM-8 | |
| Micro-Dilators | Synovis | FRS-15 RM-8d.1 | |
| Micro-Needledriver | Synovis | C-14 | |
| Micro-Clamp Applicator | Synovis | CAF-4 | |
| Micro-Flushing Needle | Hamilton | N/A | 10MM, 30°, 33G |
| Lactated Ringers Solution | Fisher Scientific | NC9968051 | |
| Buprenorphine | N/A | N/A | DEA Number required; Obtained from hosptial pharmacy. |
| Enrofloxacin; Baytril | Bayer Health Care | 186599 | |
| Heparin | N/A | N/A | Obtained from hosptial pharmacy |
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