title: Production of human tissue-engineered skin trilayer on a plasma-based hypodermis.
abstract: Full thickness wounds require a dermal component to achieve functional permanent skin restoration. Currently available tissue-engineered skin substitutes lack a subcutaneous fat layer that would functionally contribute some of the mechanical and thermoregulatory properties of normal skin. To generate a trilayer engineered skin equivalent, we included bone marrow mesenchymal (BM-MSC) or adipose tissue-derived (ASC) stromal cells in a human plasma hydrogel exposed to adipogenic clues for three weeks. Approximately half of the cells differentiated under these conditions into mature adipocytes that survived for two years in culture with minimal medium change. In vitro generation of bona fide fully differentiated adipocytes was assessed by leptin secretion and ultrastructurally demonstrated through semithin to ultrathin sectioning and lipid staining with osmium tetroxide. Furthermore, presence of BM-MSCs or ASCs within the subcutaneous layer contributed to the epidermal differentiation program, with more proliferating basal cells depositing basal membrane proteins and differentiating into mature keratinocytes that were able to generate a pluristratified epithelium. In conclusion, we engineered a fully differentiated human skin trilayer that could present multiple applications such as use for in vitro drug absorption tests and regenerative therapies.
authors.affiliation: Fundación Inbiomed, San Sebastián, Spain.
authors.forename: Asun
authors.initials: A
authors.lastname: Monfort
authors.forename: Mario
authors.initials: M
authors.lastname: Soriano-Navarro
authors.forename: José Manuel
authors.initials: JM
authors.lastname: García-Verdugo
authors.forename: Ander
authors.initials: A
authors.lastname: Izeta
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D050156:Adipogenesis / Q000187:drug effects; D000273:Adipose Tissue / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D020405:Dermis / Q000166:cytology; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D020738:Leptin / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010949:Plasma / Q000378:metabolism*; D012867:Skin / Q000033:anatomy & histology* / Q000187:drug effects; D040521:Subcutaneous Tissue / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
chemical_list: D020738:Leptin; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate