title: Characterization of pigmented dermo-epidermal skin substitutes in a long-term in vivo assay.
abstract: In our laboratory, we have been using human pigmented dermo-epidermal skin substitutes for short-term experiments since several years. Little is known, however, about the long-term biology of such constructs after transplantation. We constructed human, melanocyte-containing dermo-epidermal skin substitutes of different (light and dark) pigmentation types and studied them in a long-term animal experiment. Developmental and maturational stages of the epidermal and dermal compartment as well as signs of homoeostasis were analysed 15 weeks after transplantation. Keratinocytes, melanocytes and fibroblasts from human skin biopsies were isolated and assembled into dermo-epidermal skin substitutes. These were transplanted onto immuno-incompetent rats and investigated 15 weeks after transplantation. Chromameter evaluation showed a consistent skin colour between 3 and 4 months after transplantation. Melanocytes resided in the epidermal basal layer in physiological numbers and melanin accumulated in keratinocytes in a supranuclear position. Skin substitutes showed a mature epidermis in a homoeostatic state and the presence of dermal components such as Fibrillin and Tropoelastin suggested advanced maturation. Overall, pigmented dermo-epidermal skin substitutes show a promising development towards achieving near-normal skin characteristics and epidermal and dermal tissue homoeostasis. In particular, melanocytes function correctly over several months whilst remaining in a physiological, epidermal position and yield a pigmentation resembling original donor skin colour.
authors.affiliation: Tissue Biology Research Unit, University Children's Hospital Zurich, Zurich, Switzerland; Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland; Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: Sophie
authors.initials: S
authors.lastname: Böttcher-Haberzeth
authors.forename: Thomas
authors.initials: T
authors.lastname: Biedermann
authors.forename: Agnieszka S
authors.initials: AS
authors.lastname: Klar
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Widmer
authors.forename: Kathrin
authors.initials: K
authors.lastname: Neuhaus
authors.forename: Clemens
authors.initials: C
authors.lastname: Schiestl
authors.forename: Martin
authors.initials: M
authors.lastname: Meuli
authors.forename: Ernst
authors.initials: E
authors.lastname: Reichmann
mesh_terms: D000818:Animals; D001706:Biopsy; D017690:Cell Transplantation; D000071837:Fibrillins; D005347:Fibroblasts / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D008544:Melanocytes / Q000166:cytology* / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism; D010858:Pigmentation; D051381:Rats; D012867:Skin / Q000378:metabolism* / Q000473:pathology; D012880:Skin Pigmentation; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*; D014333:Tropoelastin / Q000378:metabolism
chemical_list: D000071837:Fibrillins; D008840:Microfilament Proteins; D014333:Tropoelastin
keywords: cultured human pigmented skin substitute; homoeostasis; melanocytes; tissue engineering