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Long-term expression pattern of melanocyte markers in light- and dark-pigmented dermo-epidermal cultured human skin substitutes

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Abstract

Purpose

Transplantation of pigmented tissue-engineered human autologous skin substitutes represents a promising procedure to cover skin defects. We have already demonstrated that we can restore the patient’s native light or dark skin color by adding melanocytes to our dermo-epidermal skin analogs. In this long-term study, we investigated if melanocytes in our skin substitutes continue to express markers as BCL2, SOX9, and MITF, known to be involved in survival, differentiation, and function of melanocytes.

Methods

Human epidermal melanocytes and keratinocytes, as well as dermal fibroblasts from light- and dark-pigmented skin biopsies were isolated and cultured. Bovine collagen hydrogels containing fibroblasts were prepared, and melanocytes and keratinocytes were seeded in a 1:5 ratio onto the gels. Pigmented dermo-epidermal skin substitutes were transplanted onto full-thickness wounds of immuno-incompetent rats and analyzed for the expression of melanocyte markers after 15 weeks.

Results

Employing immunofluorescence staining techniques, we observed that our light and dark dermo-epidermal skin substitutes expressed the same typical melanocyte markers including BCL2, SOX9, and MITF 15 weeks after transplantation as normal human light and dark skin.

Conclusions

These data suggest that, even in the long run, our light and dark dermo-epidermal tissue-engineered skin substitutes contain melanocytes that display a characteristic expression pattern as seen in normal pigmented human skin. These findings have crucial clinical implications as such grafts transplanted onto patients should warrant physiological numbers, distribution, and function of melanocytes.

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Acknowledgments

We thank Ines Kleiber-Schaaf (Department of Dermatology, University Hospital, Zurich) for precious help in the preparation and analysis of the probes for transmission electron microscopy and histology. This work was financially supported by the EU-FP7 project EuroSkinGraft (FP7/2007-2013: grant agreement no 279024), by the EU-FP7 (MultiTERM, grant agreement no 238551) and the Clinical Research Priority Programs (KFSP: From basic research to the clinic: Novel tissue engineered skin grafts for Zurich) of the Faculty of Medicine of the University of Zurich. We are particularly grateful to the Fondation Gaydoul and the sponsors of “DonaTissue” (Thérèse Meier and Robert Zingg) for their generous financial support and interest in our work.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Tissue Biology Research Unit, University Children’s Hospital Zurich, Zurich, Switzerland

    Thomas Biedermann, Agnieszka S. Klar, Sophie Böttcher-Haberzeth, Teresa Michalczyk & Ernst Reichmann

  2. Department of Surgery, University Children’s Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland

    Clemens Schiestl & Martin Meuli

  3. Children’s Research Center, University Children’s Hospital Zurich, Zurich, Switzerland

    Thomas Biedermann, Agnieszka S. Klar, Sophie Böttcher-Haberzeth, Teresa Michalczyk, Clemens Schiestl, Ernst Reichmann & Martin Meuli

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  1. Thomas Biedermann
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  2. Agnieszka S. Klar
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  3. Sophie Böttcher-Haberzeth
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  5. Clemens Schiestl
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  7. Martin Meuli
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Corresponding author

Correspondence to Martin Meuli.

Additional information

T. Biedermann and A. S. Klar contributed equally.

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Biedermann, T., Klar, A.S., Böttcher-Haberzeth, S. et al. Long-term expression pattern of melanocyte markers in light- and dark-pigmented dermo-epidermal cultured human skin substitutes. Pediatr Surg Int 31, 69–76 (2015). https://doi.org/10.1007/s00383-014-3622-7

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