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. 2025:2922:135-142.
doi: 10.1007/978-1-0716-4510-9_10.

Engineering a Multilayered Skin Substitute with Keratinocytes, Fibroblasts, Adipose Derived Stem Cells and Adipocytes

Maike Keck  1   2 , Alfred Gugerell  3 , Johanna Kober  4
Affiliations

Engineering a Multilayered Skin Substitute with Keratinocytes, Fibroblasts, Adipose Derived Stem Cells and Adipocytes

Maike Keck et al. Methods Mol Biol. 2025.

Abstract

A variety of skin substitutes that restore epidermal and dermal structures are currently available on the market. While the main focus in research and clinical application lies in dermal and epidermal substitutes, the development of a subcutaneous replacement, the hypodermis, is often neglected. This chapter describes the use of fibrin sealant as a hydrogel scaffold to generate a three dimensional skin substitute. For the hypodermal layer, adipose-derived stem cells (ASCs) and mature adipocytes are seeded within a fibrin hydrogel. On top, another fibrin clot with incorporated fibroblasts is placed for the construction of the dermal layer. Keratinocytes are added on top of the two-layered construct to form the epidermal layer. The three layered construct is cultivated for up to 3 weeks with keratinocytes being exposed to air according to the air-liquid interface cultivation model.

Keywords: Adipocytes; Adipose derived stem cells; Fibroblasts; Keratinocytes; Regenerative medicine; Three layered skin substitute; Tissue engineering.

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References

    1. Peterbauer-Scherb A, Danzer M, Gabriel C et al (2012) In vitro adipogenesis of adipose-derived stem cells in 3D fibrin matrix of low component concentration. J Tissue Eng Regen Med 6(6):434–442 - DOI - PubMed
    1. Siegel KR, Clevenger TN, Clegg DO et al (2018) Adipose stem cells incorporated in fibrin clot modulate expression of growth factors. Arthroscopy 34(2):581–591. https://doi.org/10.1016/j.arthro.2017.08.250 - DOI - PubMed
    1. Kober J, Gugerell A, Schmid M et al (2015) Generation of a fibrin based three-layered skin substitute. Biomed Res Int. https://doi.org/10.1155/2015/170427
    1. Keck M, Kober J, Riedl O et al (2014) Power assisted liposuction to obtain adipose-derived stem cells: impact on viability and differentiation to adipocytes in comparison to manual aspiration. J Plast Reconstr Aesthet Surg 67(1):E1–E8 - DOI - PubMed
    1. Mildmay-White A, Khan W (2017) Cell surface markers on adipose-derived stem cells: a systematic review. Curr Stem Cell Res Ther 12(6):484–492 - DOI - PubMed

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