Allogeneic skin transplantation is a standard model to assay host T cell responses to MHC-disparate donor antigens. This video-article provides a visual tutorial of each step involved in performing a BALB/c-->C57BL/6 skin transplant.
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In This Article
Summary
Abstract
As one of the most stringent and least technically challenging models, skin transplantation is a standard method to assay host T cell responses to MHC-disparate donor antigens. The aim of this video-article is to provide the viewer with a step-by-step visual demonstration of skin transplantation using the mouse model. The protocol is divided into 5 main components: 1) harvesting donor skin; 2) preparing recipient for transplant; 3) skin transplant; 4) bandage removal and monitoring graft rejection; 5) helpful hints. Once proficient, the procedure itself should take <10 min to perform.
Protocol
Prior to the initiation of experiment:
- Obtain IACUC approval
- Sterilize surgical instruments and Q-tips
I. Harvest Donor Tissue
- Euthanize donor mouse (in accordance with institution's SOP)
- Wet hair back from base of ear with alcohol
- Harvest donor skin by cutting ear off at base
- Place tissue in plate on ice
- Split ear: pinch ventral and dorsal sides of tissue at base of ear with rat-tooth forceps; gently pull apart tissue; float dorsal tissue, internal surface down, in PBS on ice; discard collagenous ventral flap
II. Prepare Recipient for Tr....
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Discussion
As demonstrated in this video-article, the skin transplant model is a quick and easy method for monitoring allogeneic T cell responses. Allogeneic skin transplantation has a wide range of applications and has been utilized to assay the efficiency of a variety of immunosuppressive agents and tolerance induction protocols, as well as to investigate the biological effects of various blocking antibodies.
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Disclosures
The authors have nothing to disclose.
Acknowledgements
NIH Immunology Training Grant T32 AI 60573 (KRG) and NIH grant GM 41514 (MDC)
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Micro-dissecting forceps (curved) | Sigma-Aldrich | F4142 | |
| Rat-tooth forceps (1x2 teeth, straight) | Fine Science Tools | 11053-10 | n=2 |
| Strabismus scissors (blunt end) | Fine Science Tools | 14074-11 | |
| Iris forceps (curved) | Fine Science Tools | 11065-07 | |
| Spring scissors (angled) | Fine Science Tools | 15006-09 | |
| Halsey micro needle holder | Fine Science Tools | 12500-12 | |
| 10 cm plates | Corning | 430167 | |
| PBS | Cellgro | 21-040-CV | |
| Bupernex (Buprenorphine hydrochloride) | Reckitt Benckiser | 517201 | |
| Insulin syringes (29 Ga) | Monoject | 8881 600145 | |
| Q-tips | Solon Manufacturing Company | 362 | |
| Single edge razor blades (#9) | Smith & Nephew Inc. | 67-0238 | |
| Silk suture (2-0; PS-2) | Ethicon Inc. | 583H |
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