Integration of Mature Adipocytes to Build-Up a Functional Three-Layered Full-Skin Equivalent
| Autor: | Birgit Huber, Sandra Gehrke, Marc Winnefeld, Antonia Link, Kirstin Linke, Petra J. Kluger |
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| Přispěvatelé: | Publica |
| Rok vydání: | 2016 |
| Předmět: |
Keratinocytes
0301 basic medicine Cellular differentiation Biomedical Engineering Mature adipocytes Medicine (miscellaneous) chemistry.chemical_element Bioengineering Calcium Article 03 medical and health sciences chemistry.chemical_compound Equivalent Adipocyte Adipocytes Humans Skin equivalent Cells Cultured Skin Skin Artificial Tissue Engineering integumentary system Cell Differentiation Fibroblasts Coculture Techniques In vitro Cell biology 030104 developmental biology chemistry Toxicity Cytokines Biomedical engineering |
| Zdroj: | Tissue Engineering. Part C, Methods |
| ISSN: | 1937-3392 1937-3384 |
| DOI: | 10.1089/ten.tec.2016.0141 |
| Popis: | Large, deep full-thickness skin wounds from high-graded burns or trauma are not able to reepithelialize sufficiently, resulting in scar formation, mobility limitations, and cosmetic deformities. In this study, in vitro-constructed tissue replacements are needed. Furthermore, such full-skin equivalents would be helpful as in vivo-like test systems for toxicity, cosmetic, and pharmaceutical testing. Up to date, no skin equivalent is available containing the underlying subcutaneous fatty tissue. In this study, we composed a full-skin equivalent and evaluated three different media for the coculture of mature adipocytes, fibroblasts, and keratinocytes. Therefore, adipocyte medium was supplemented with ascorbyl-2-phosphate and calcium chloride, which are important for successful epidermal stratification (Air medium). This medium was further supplemented with two commercially available factor combinations often used for the in vitro culture of keratinocytes (Air-HKGS and Air-KGM medium). We showed that in all media, keratinocytes differentiated successfully to build a stratified epidermal layer and expressed cytokeratin 10 and 14. Perilipin A-positive adipocytes could be found in all tissue models for up to 14 days, whereas adipocytes in the Air-HKGS and Air-KGM medium seemed to be smaller. Adipocytes in all tissue models were able to release adipocyte-specific factors, whereas the supplementation of keratinocyte-specific factors had a slightly negative effect on adipocyte functionality. The permeability of the epidermis of all models was comparable since they were able to withstand a deep penetration of cytotoxic Triton X in the same manner. Taken together, we were able to compose functional three-layered full-skin equivalents by using the Air medium. |
| Databáze: | OpenAIRE |
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