A Technique for Human Skin Grafting in a Mouse Model

Abstract

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.

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 choice of graft size.

1. Maintain the skin sample at cold temperatures (on ice; 4°C) before preparation and grafting. Keep the sample moist....

Results

Figure 1: Timeline of the protocol adopted for the xenograft study.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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