title: Type IV collagen aggregates promote keratinocyte proliferation and formation of epidermal layer in human skin equivalents.
abstract: Type IV collagen isolated from lens capsule without enzymatic treatment is known to form a gel under physiological condition and influences cellular activities. In case of human keratinocytes, the suppression of proliferation on reconstituted type IV collagen gels was reported in monolayer culture. In this study, we examined effects of type IV collagen isolated from porcine lens capsule on epidermal formation in human skin equivalents (HSEs). Type IV collagen aggregates were prepared under the culture condition and the aggregates suppressed keratinocyte proliferation in monolayer culture as well as the culture on the gels. In HSEs, type IV collagen aggregates were reconstituted on the surface of contracted collagen gels containing human dermal fibroblasts and the keratinocytes were then cultured on the aggregates for 14 days. Interestingly, in HSEs with type IV collagen aggregates, the BrdU-positive keratinocytes were increased and the thickness of the epidermal layer was around twice than that of control culture. Epidermal differentiation markers were expressed in the upper layer of the epidermis and the defined deposition of human basement membrane components were increased at the dermal-epidermal junction. These results indicate that the type IV collagen aggregates stimulate the proliferation of basal keratinocytes and improve the stratification of epidermal layers in HSEs.
authors.affiliation: Scleroprotein Research Institute, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Matsuura-Hachiya
authors.affiliation: Scleroprotein Research Institute, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
authors.forename: Koji Y
authors.initials: KY
authors.lastname: Arai
authors.affiliation: Pharmaceutical and Healthcare Research Laboratories, Fuji Film Co., Kaisei, Kanagawa, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Muraguchi
authors.affiliation: Pharmaceutical and Healthcare Research Laboratories, Fuji Film Co., Kaisei, Kanagawa, Japan.
authors.forename: Tasuku
authors.initials: T
authors.lastname: Sasaki
authors.affiliation: Scleroprotein Research Institute, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
authors.forename: Toshio
authors.identifier: 0000-0002-3459-2636
authors.initials: T
authors.lastname: Nishiyama
mesh_terms: D001485:Basement Membrane; D049109:Cell Proliferation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000502:physiology*; D046508:Culture Techniques; D004817:Epidermis; D006801:Humans; D015603:Keratinocytes / Q000502:physiology*
chemical_list: D024141:Collagen Type IV
keywords: human skin equivalent; keratinocyte; proliferation; type IV collagen