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The mouse excisional wound splinting model, including applications for stem cell transplantation

Nature Protocols volume 8, pages 302–309 (2013)Cite this article

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Abstract

The mouse excisional wound healing model has been used extensively to study wound healing and cutaneous regeneration. However, as mouse skin is mobile, contraction accounts for a large part of wound closure. In the mouse excisional wound splinting model, a splinting ring tightly adheres to the skin around the wound, preventing local skin contraction. The wound therefore heals through granulation and re-epithelialization, a process similar to that occurring in humans. The model, which takes 2–4 weeks to carry out, can be used to study the effects of stem cells on cutaneous repair or regeneration. In this protocol, we also describe how to implant stem cells onto the wound bed in Matrigel and/or into the surrounding tissue through injection. Serial wound tissue samples at different time points can be harvested to monitor the engraftment and the effects of stem cells in angiogenesis and wound healing.

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Figure 1: Splinting wounds versus conventional wounds.
Figure 2: An illustration of excisional wound splinting model and stem cell transplantation.
Figure 3: Engraftment of mesenchymal stem cells into the wound.
Figure 4: FACS analyses of wound digests for GFP-positive mesenchymal stem cells.
Figure 5: Neovasculature in wounds.

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Acknowledgements

This work is supported by grants from the Natural Science Foundation of China (nos. 30971496 and U1032003), from Shenzhen (JC201005280597A) to Y.W., and by the Firefighters' Burn Trust Fund of The University of Alberta to E.E.T.

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Authors and Affiliations

  1. School of Life Sciences, Tsinghua University, Beijing, China

    Xusheng Wang & Jianfeng Ge

  2. The Shenzhen Key Laboratory of Health Sciences and Technology, Graduate School at Shenzhen, Tsinghua University, Shenzhen, China

    Xusheng Wang, Jianfeng Ge & Yaojiong Wu

  3. Department of Surgery, Wound Healing Research Group, University of Alberta, Edmonton, Alberta, Canada

    Edward E Tredget

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  1. Xusheng Wang
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  2. Jianfeng Ge
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  3. Edward E Tredget
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  4. Yaojiong Wu
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Contributions

E.E.T. and Y.W. designed the studies; X.W., J.G. and Y.W. performed the experiments and analyzed the data; and Y.W. wrote the manuscript.

Corresponding author

Correspondence to Yaojiong Wu.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Video 1

Creation of murine excisional wound splinting model and transplantation of stem cells. All animal experiments were performed with the approval of the Animal Ethics Committee of Tsinghua University. Splints are prepared from a silicon sheet. After anesthesia, two equal sized full-thickness wounds are created on the depilated dorsal skin of a Balb/C mouse with a biopsy punch. Stem cells are injected into the dermis around the wound. Spread an instant-bonding adhesive on one side of a splint and carefully place the splint around the wound so that the wound is centered within the splint. Secure the splint to the skin with interrupted sutures. Apply stem cells in Matrigel onto the wound bed. Cover the wounds and splints with sterile transparent dressing Tegaderm. Dress the wounds with self-adhering elastic bandage. (MOV 15465 kb)

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Wang, X., Ge, J., Tredget, E. et al. The mouse excisional wound splinting model, including applications for stem cell transplantation. Nat Protoc 8, 302–309 (2013). https://doi.org/10.1038/nprot.2013.002

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