| Project/Area Number |
16380227
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
NISHIYAMA Toshio Tokyo University of Agriculture and Technology, Faculty of Agriculture, Professor, 農学部, 教授 (60372455)
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| Co-Investigator(Kenkyū-buntansha) |
ARAI Katsuhiko Tokyo University of Agriculture and Technology, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (60175940)
岸本 治郎 資生堂ライフサイエンス研究センター, 副主任研究員
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,500,000 (Direct Cost: ¥15,500,000)
Fiscal Year 2006: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 2005: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2004: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | skin equivalent / collagen / basement membrane / epidermal differentiation / fibroblast / keratinocyte / matrix metalloproteinase / mechanical stress / 前線維芽細胞 / MMP / 細胞外マトリックス / 培養人口皮膚 / ラミニン / 繊維芽細胞 |
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| Research Abstract |
In vitro models of human skin utilizing keratinocytes grown upon fibroblast-contracted collagen gels have been known to demonstrate similar behavior to the cells in vivo. The purpose of our study is to provide suitable bioassay models for investigating the structure and function of epidermal or dermal tissue and to determine the dermal-epidermal interactions required for keratinocyte function, basement membrane formation and hair follicle formation utilizing the three-dimensional organotypic cultures such as skin-equivalents (SEs). Despite production of the epidermal basement membrane (BM) components, BM was rarely observed in SEs. Analyses through SEs have revealed that matrix metalloproteinases (MMPs) and plasmin are involved in BM formation. To elucidate the affect of these proteinases, we examined effects of inhibitors for MMPs and serine proteinase on the BM formation and the epidermal structure and differentiation in the SEs. By addition of both inhibitors the epidermis was well
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organized with multilayered keratinocytes consisting of basal, spinous, granular, and cornified layers. The epidermal differentiation markers were expressed in the upper layer of the epidermis and the sharply defined depositions specific for BM components were observed at the dermal-epidermal junction. Ultrastructurally, the extended and continuous lamina densa with approximately 30 nm thick was observed underneath the basal keratinocytes. These results suggest that the assembly of BM under the basal keratinocytes and the differentiation of epidermis are extensively promoted under the influence of inhibitors for MMPs and serine proteinases. In attempt on hair follicle formation by using this culture system, mouse dermal papilla cells were cultured at the dermal-epidermal interface in SEs. Cellular masses including the dermal papilla cells were frequently observed in the upper part of the dermis. Further studies are needed to determine whether the cellular masses are similar to primitive hair follicles. Less
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