title: 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals.
abstract: Stable protein microcrystals called polyhedra are produced by certain insect viruses. Cytokines, such as fibroblast growth factors (FGFs), can be immobilized within polyhedra. Here, we investigated three-dimensional (3D) co-cultures of keratinocytes and melanocytes on collagen gel containing FGF-2 and FGF-7 polyhedra. Melanocytes were observed to reside at the base of the 3D cell culture and melanin was also typically observed in the lower layer. The 3D cell culture model with FGF-2 and FGF-7 polyhedra was a useful in vitro model of the epidermis due to effective melanogenesis, proliferation and differentiation of keratinocytes. FGF-7 polyhedra showed a potent cytoprotective effect when keratinocytes were treated with menadione, which is a generator of reactive oxygen species. The cytoprotective effect was activated by the inositol triphosphate kinase-Akt pathway leading to upregulation of the antioxidant enzymes superoxide dismutase and peroxiredoxin 6.
authors: Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan.
authors: Junji
authors: J
authors: Shimabukuro
authors: Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan.
authors: Ayako
authors: A
authors: Yamaoka
authors: Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan.
authors: Ken-Ichi
authors: K
authors: Murata
authors: Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan; Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto, Japan.
authors: Eiji
authors: E
authors: Kotani
authors: Venture Laboratory, Kyoto Institute of Technology, Kyoto, Japan.
authors: Tomoko
authors: T
authors: Hirano
authors: Functional Genomics Group, COMB, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa, Japan.
authors: Yumiko
authors: Y
authors: Nakajima
authors: Division of Oral Surgery, Yokohama Clinical Education Center of Kanagawa Dental University, Yokohama, Japan.
authors: Goichi
authors: G
authors: Matsumoto
authors: Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan; Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto, Japan. Electronic address: hmori@kit.ac.jp.
authors: Hajime
authors: H
authors: Mori
mesh_terms: D018920:Coculture Techniques; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000494:pharmacology*; D007304:Insect Viruses; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects*; D008544:Melanocytes / Q000166:cytology / Q000187:drug effects*; D020011:Protective Agents / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D014764:Viral Proteins / Q000737:chemistry / Q000494:pharmacology
chemical_list: D020011:Protective Agents; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D014764:Viral Proteins; D016222:Fibroblast Growth Factor 2
keywords: 3D cell culture; Cytoprotection; FGF; Keratinocytes; Melanocytes; Polyhedra