Orthotopic Hind Limb Transplantation in the Mouse

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.

Vascularized composite allotransplantation combined with costimulation blockade induces mixed chimerism and reveals intrinsic tolerogenic potential

“…CoB promotes VCA survival. To test the effect of CoB on VCA survival, we used our recently reported murine orthotopic hind limb transplant model (15) performed across a full MHC barrier: from Balb/C (H-2 d ) into C57BL/6 (H-2 b ) ( Figure 1, A-C). Treatment with CTLA4-Ig alone showed prolonged VCA survival (median survival time [MST] 17 days) as compared with untreated controls (MST 8 days; P < 0.01).…”

“…Orthotopic hind limb transplantation and full-thickness skin transplantation. Balb/C hind limbs were transplanted into C57BL/6 recipients using a modified version of the technique described by Furtmüller et al (15). Using the previously described 4-grade rejection scale, we defined Grade 3 (skin epidermolysis) as the endpoint in this study.…”

Split Tolerance in a Murine Model of Heterotopic En Bloc Chest Wall Transplantation

“…Clinically, this rejection profile is similar to previous results of hind limb allograft rejection in 4 distinct rejection stages—beginning with skin loss. 16 In allogeneic recipients under costimulation blockade treatment, skin rejection was observed after 13 ± 1 days. However, we observed long-term survival of all other allograft components, exceeding 130 days (Fig.…”

“…Multiple murine transplant models for either organ transplantation or VCA are available. 16 , 17 In contrast, models are needed for an investigation of combined SOT and VCA. The major advantage of the en bloc chest wall, thymus, and heart transplant murine model is that it includes both types of transplant and also a thymic component that has the potential to support T-cell development.…”

Improved Cuff Technique and Intraoperative Detection of Vascular Complications for Hind Limb Transplantation in Mice

“… 19 The technique was also adapted to a mouse model. 20 , 21 However, the mounting of a mouse femoral artery with a microtube is time-consuming due to the diminutive size of the femoral artery. Even though more collateral vessels required ligation by using the AA, the donor surgery time was much shorter, 52 ± 10 minutes, than previously published methods using the femoral artery, 100 ± 12 minutes.…”