Formation of keratinocyte multilayers on filters under airlifted or submerged culture conditions in medium containing calcium, ascorbic acid, and keratinocyte growth factor.

• Material Type: 博士論文
• Title: Formation of keratinocyte multilayers on filters under airlifted or submerged culture conditions in medium containing calcium, ascorbic acid, and keratinocyte growth factor.
• Author/Editor: Seo, Akira; Kitagawa, Norio; Matsuura, Takashi; Sato, Hironobu; Inai, Tetsuichiro
• Author Heading: Seo, Akira; Kitagawa, Norio; Matsuura, Takashi; Sato, Hironobu; Inai, Tetsuichiro
• Publication Date: 2017-03-11
• Publication Date (W3CDTF): 2017-03-11
• Alternative Title: Formation of keratinocyte multilayers on filters under airlifted or submerged culture conditions in medium containing calcium, ascorbic acid, and keratinocyte growth factor.
• Degree grantor/type: 福岡歯科大学
• Date Granted: 2017-03-11
• Date Granted (W3CDTF): 2017-03-11
• Dissertation Number: 甲第283号
• Degree Type: 博士（歯学）
• Conferring No. (Dissertation): 甲第283号
• Text Language Code: eng
• Subject Heading: Adult; Ascorbic Acid; Calcium; Cells, Cultured; Culture Media; Fibroblast Growth Factor 7; Humans; Keratinocytes; Cytokeratins; Integrin β4; Involucrin; Organotypic culture
• Target Audience: 一般
• Note (General): Three-dimensional (3D) cell culture is a powerful in vitro technique to study the stratification and differentiation of keratinocytes. However, culture conditions, including culture media, supplements, and scaffolds (e.g., collagen gels with or without fibroblasts), can vary considerably. Here, we evaluated the roles of calcium, L-ascorbic acid phosphate magnesium salt n-hydrate (APM), and keratinocyte growth factor (KGF) in a chemically defined medium, EpiLife, in 3D cultures of primary human epidermal keratinocytes directly plated on polycarbonate filter inserts under airlifted or submerged conditions. Eight culture media containing various combinations of these three supplements were examined. Calcium was necessary for the stratification and differentiation of keratinocytes based on the localization of keratins and involucrin. However, the localization patterns of keratins and integrin β4 were partially disrupted and Ki67-positive basal cells almost disappeared 3weeks after airlift. The addition of KGF, but not APM, prevented these changes. Further addition of APM markedly improved the tissue architecture, including basal cell morphology and the appearance of keratohyalin granules and localized involucrin in the upper suprabasal cells, even after 1week. Although the submerged culture also formed cornified epithelium-like multilayers, involucrin was localized in the cornified layer, where nuclei were often found. Based on these results, it is most effective to culture keratinocytes at the air-liquid interface in EpiLife medium supplemented with calcium, APM, and KGF to form well-organized and orthokeratinized multilayers as skin analogues.; SpringerLink
• Persistent ID (NDL): info:ndljp/pid/10951995; https://dl.ndl.go.jp/pid/10951995
• Collection: 障害者向け資料
• Collection (Materials For Handicapped People:1): テキストデータ
• Collection (particular): 国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文; https://dl.ndl.go.jp/collections/A00014
• Acquisition Basis: 博士論文（自動収集）
• Date Accepted (W3CDTF): 2017-09-02T15:28:24+09:00
• Date Created (W3CDTF): 2017-08-01
• Format (IMT): application/pdf
• Access Restrictions: 国立国会図書館内限定公開
• Service for the Digitized Contents Transmission Service: 図書館・個人送信対象外
• Availability of remote photoduplication service: 可
• Original Material (URI): https://link.springer.com/article/10.1007%2Fs00418-016-1472-1
• Periodical Title (URI): https://fdc.repo.nii.ac.jp/records/76
• Data Provider (Database): 国立国会図書館 : 国立国会図書館デジタルコレクション; https://dl.ndl.go.jp

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