【博士論文】学術データベース

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

著者

書誌事項

タイトル

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

著者名

Seo Akira

著者名

Kitagawa Norio

著者名

Matsuura Takashi

著者名

Sato Hironobu

著者名

Inai Tetsuichiro

学位授与大学

福岡歯科大学 (大学ID:2285) (CAT機関ID:KI004437)

取得学位

博士(歯学)

学位授与番号

甲第283号

学位授与年月日

2017-03-11

注記・抄録

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 3 weeks 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 1 week. 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.

2016年度

https://link.springer.com/article/10.1007%2Fs00418-016-1472-1

キーワード

Calcium, Adult, Ascorbic Acid, Cells, Cultured, Culture Media, Fibroblast Growth Factor 7, Humans, Keratinocytes, Cytokeratins, Integrin β4, Involucrin, Organotypic culture

各種コード

NII論文ID(NAID)

500001033449

NII著者ID(NRID)
  • 8000001142380
  • 8000001142381
  • 8000001142382
  • 8000001142383
  • 8000001142384
本文言語コード

eng

データ提供元

機関リポジトリ / NDLデジタルコレクション

外部リンク

博士論文 / 福岡歯科大学 / 歯学

博士論文 / 福岡歯科大学

博士論文 / 歯学

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