Murine Full-thickness Skin Transplantation

Abstract

Murine full-thickness skin transplantation is a well-established in vivo model to study alloimmune response and graft rejection. Despite its limited application to humans, skin transplantation in mice has been widely employed for transplantation research. The procedure is easy to learn and perform, and it does not require delicate microsurgical techniques nor extensive training. Moreover, graft rejection in this model occurs in a very reproducible immunological reaction and is easily monitored by direct inspection and palpation. In addition, secondary skin transplantation with donor-matched or third-party skin grafts can be performed on more complex transplant models as an alternative and uncomplicated method to assess donor-specific tolerance. The complications are low and are in general limited to anesthesia overdose or respiratory distress after the procedure. Graft failure, on the other hand, occurs commonly as a result of poor preparation of the graft, incorrect positioning in the graft bed, or inappropriate placement of the bandage. In this article, we present a protocol for full-thickness skin transplantation in mice and describe the important steps necessary for a successful procedure.

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

“…Full-thickness trunk skin from Balb/C (donor-matched) or FVB/N (third-party) mice was transplanted onto the lateral thoracic wall of C57BL/6 recipients and secured with 6.0 Ethilon sutures (44). Skin graft survival was monitored daily.…”

Recombinant human ADAMTS13 treatment and anti-NET strategies enhance skin allograft survival in mice

“…Their dorsal skin was rapidly shaved, sterilized alternatively with 70% ethanol wipes and betadine 3 times, and harvested. Connective tissue, fat tissue, and panniculus carnosus were removed from the donor’s skin, which was then cut into 1 × 1‐cm grafts and kept in phosphate‐buffered saline (PBS) on ice . After graft preparation, the dorsal skin of the recipient mice was shaved and sterilized.…”

“…Acute graft rejection began with swelling and erythema, followed by graft desiccation, shrinkage, and scab formation. Complete rejection was defined as complete hardening of the graft with loss of hair …”

“…Connective tissue, fat tissue, and panniculus carnosus were removed from the donor's skin, which was then cut into 1 × 1-cm grafts and kept in phosphate-buffered saline (PBS) on ice. 23 After graft preparation, the dorsal skin of the recipient mice was shaved and sterilized. With proper anesthesia using isoflurane inhalation, a piece of full-thickness skin (1 × 1 cm) was excised from the back of the recipient mice, followed by suturing of the allograft to the wound.…”

“…Human skin and allograft specimens, as well as allografts of vehicle-or rhADAMTS13treated mice harvested on day 3, were cryosectioned into 10-μm sections. The sections were postfixed in zinc fixative (100 mmol/L Tris-HCl, 37 mmol/L zinc chloride,23 mmol/L zinc acetate, 3.2 mmol/L calcium acetate), permeabilized with 0.1% Triton-X and 0.1% sodium citrate for 10 minutes at 4°C, blocked with 3% bovine serum albumin (BSA) or 2.5% BSA/5% donkey serum in PBS, and incubated with combinations of primary antibodies against H3Cit (Abcam, Cambridge, MA; ab5103, 1:1000), H4Cit (Millipore, Burlington, MA; Cat. no.…”