Source: Moss, M. I. et al., Xenograft Skin Model to Manipulate Human Immune Responses In Vivo. J. Vis. Exp. (2022)
This video demonstrates a method for grafting human skin onto an immunodeficient mouse with a suppressed neutrophil response. The method involves applying human split-thickness skin to the mouse's graft bed, sealing it, and covering it for recovery. Eventually, the human skin adheres to the mouse's skin, establishing vascularization and circulation, resulting in a successful skin xenograft.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Processing of donor human skin sample
NOTE: The human skin sample used in this transplant was a large sample collected from the abdomen of a healthy patient. The sample must be at least 15 cm x 7.5 cm. Size limitations may affect the number of mice for which skin is available and the choice of graft size.
2. Pre-surgery conditioning and preparation
3. Transplantation procedure
Figure 1: Timeline of the protocol adopted for the xenograft study.
Figure 2: Method for bandaging mouse after surgery. The mouse is wrapped tightly in petrolatum gauze and transparent film dressing while under anesthesia. (A) Petrolatum gauze is placed over an area slightly larger than the surgical wound. (B) The transparent film dressing is prepared: a long strip is cut slightly wider than the petrolatum gauze. The backing covering the adhesive side of the transparent film dressing is partially removed. (C) The adhesive side of the film is firmly pressed against the back of the mouse, leaving an inch of space on the front end of the film. (D) The mouse is turned on its back. The overhanging front portion of the film is pressed onto the mouse, and the backing is removed. (E) The mouse is tightly wrapped in the film, and the remaining support is removed as the mouse is wrapped. (F) The mouse is successfully wrapped in the dressing, and its arms and legs are unconstrained.
The authors have nothing to disclose.
10% Neutral Buffered Formalin | Fisher | SF100-20 | Fixative for histology |
3M Vetbond Tissue Adhesive | 3M | 1469SB | Surgical glue |
Anti-GR1 clone RB6-8C5 | BioXcell | BE0075 | Anti-rejection |
Autoclave pouches | VWR | 89140-800 | For autoclaving tools and paper towels |
Buprenex 0.3 mg/mL | Covetrus | 59122 | Analgesia |
Carprofen 50 mg/mL | Zoetis | NADA # 141-199 | Analgesia |
D42 Dermatome blade | Humeca | 5.D42BL10 | Dermatome (1 blade per sample) |
Dermatome D42 | Humeca | 4.D42 | Dermatome |
Disposable Scalpel | Bard-Parker | 371610 | skin preparation |
Dissecting T-Pins; 1-1/2 inch, 1000/CS 1.5 | Cole-Parmer | UX-10915-03 | To pin skin specimen for dermatome |
Dissection scissors | medicon | 02.04.10 | Sample preparation and mouse dissection |
DNAse | Sigma-Aldrich | DN25-1G | Skin digestion |
Electric clippers | Kent | CL8787-KIT | Hair removal |
Forceps | medicon | 07.60.07 | Sample preparation and mouse dissection |
Gauze | Fisherbrand | 22-362-178 | Sample preparation |
Heating lamp | Morganville Scientific | HL0100 | Post-surgical care |
Heating pads 4" x 10" | Pristech | 20415 | Surgical heat supply |
Insulin 1cc 12.7 mm syringes | BD | 329410 | Drug administration |
Isoflurane | United States Pharmacopeia (USP) | NDC 66794-013-25 | Anesthesia |
Isoflurane machine | VetEquip | 911103 | Anesthesia |
Nair for Men | Nair | 10022600588556 | Hair removal |
Neomycin and Polymyxin Bisulfates and Bacitracin Zinc Ophthalmic ointment | Dechra | NDC 17478-235-35 | Eye ointment to prevent drying |
NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice | The Jackson Laboratory | 5557 | Mice |
Paper towels | Kleenex | 100848 | May be autoclaved for sterile surfaces |
Parafilm | Fisher Scientific | 13-374-12 | Semitransparent sealing film |
Petri Dish 150 mm | Corning | 430597 | Sample storage |
Plastic Wrap | Fisherbrand | 22-305-654 | Site preparation |
Providone-Iodine Swab stick | PDI | S41350 | Site sterilization |
Soft-Feed and Oral Hydration (Napa Nectar) | Se Lab Group Inc | NC9066511 | For supplementing poorly recovering mice post-surgery |
Specimen Collection Cups | Fisher Scientific | 22-150-266 | Sample storage |
Sterile alcohol prep pad | Fisherbrand | 22-363-750 | Skin preparation |
Sterile PBS | Gibco | 14190-144 | Media for sample storage |
Sterile saline | Hospira | NDC 0409-4888-02 | For drug dilution |
Tegaderm Film 4" x 43/4" | 3M | 1626 | Transparent film wound dressing |
Vaseline Petrolatum Gauze 3" x 8" | Kendall | 414600 | Wound dressing |