title: Cellular artificial skin substitute produced by short period simultaneous culture of fibroblasts and keratinocytes.
abstract: We developed a novel artificial skin substitute consisting of two collagen sponge layers with different pore sizes and cross-link densities. Fibroblasts suspended in 0.5 ml Dulbecco-modified Eagle's medium (DMEM) + 10% fetal bovine serum (FBS) were seeded on the lower dermal sponge layer, then epidermal collagen sponge and 0.1 ml suspension of keratinocytes in KGM were layered in this order. After a few hours, the medium was changed to DMEM + 5% FBS. These processes were carried out in one day, and the composite layers were then cultured by the air-liquid interface culture method. Three to five days after seeding, keratinocytes had grown to about ten layers, and fibroblasts had grown three-dimensionally into the lower dermal sponge layer. This novel cellular artificial skin substitute was grafted onto nude mice and took in 4 weeks. This skin substitute has the advantage of a shorter culturing period than previously cultured skins, and may be clinically useful for grafting that is urgently required in patients with severe generalised burns.
authors.affiliation: Department of Plastic Surgery, Kyoto University, Kyoto.
authors.forename: B M
authors.initials: BM
authors.lastname: Kim
authors.forename: S
authors.initials: S
authors.lastname: Suzuki
authors.forename: Y
authors.initials: Y
authors.lastname: Nishimura
authors.forename: S C
authors.initials: SC
authors.lastname: Um
authors.forename: K
authors.initials: K
authors.lastname: Morota
authors.forename: T
authors.initials: T
authors.lastname: Maruguchi
authors.forename: Y
authors.initials: Y
authors.lastname: Ikada
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology*; D006085:Graft Survival; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D012867:Skin / Q000166:cytology; D017768:Skin, Artificial