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.
Protocol
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 choic…
Representative Results
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 …
Divulgazioni
The authors have nothing to disclose.
Materials
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