オカノ　テルオ   OKANO Teruo   岡野　光夫    所属   研究施設 研究施設    職種   特任顧問
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	A novel closed cell culture device for fabrication of corneal epithelial cell sheets.
掲載誌名	正式名：Journal of tissue engineering and regenerative medicine 略　 称：J Tissue Eng Regen Med ISSNコード：19327005/19326254
掲載区分	国外
巻・号・頁	9(11),pp.1259-1267
著者・共著者	NAKAJIMA Ryota†, KOBAYASHI Toyoshige, MORIYA Noboru, MIZUTANI Manabu, KAN Kazutoshi, NOZAKI Takayuki, SAITOH Kazuo, YAMATO Masayuki, OKANO Teruo, TAKEDA Shizu*
発行年月	2015/11
概要	Automation technology for cell sheet-based tissue engineering would need to optimize the cell sheet fabrication process, stabilize cell sheet quality and reduce biological contamination risks. Biological contamination must be avoided in clinical settings. A closed culture system provides a solution for this. In the present study, we developed a closed culture device called a cell cartridge, to be used in a closed cell culture system for fabricating corneal epithelial cell sheets. Rabbit limbal epithelial cells were cultured on the surface of a porous membrane with 3T3 feeder cells, which are separate from the epithelial cells in the cell cartridges and in the cell-culture inserts as a control. To fabricate the stratified cell sheets, five different thicknesses of the membranes which were welded to the cell cartridge, were examined. Multilayered corneal epithelial cell sheets were fabricated in cell cartridges that were welded to a 25 µm-thick gas-permeable membrane, which was similar to the results with the cell-culture inserts. However, stratification of corneal epithelial cell sheets did not occur with cell cartridges that were welded to 100-300 µm-thick gas-permeable membranes. The fabricated cell sheets were evaluated by histological analyses to examine the expression of corneal epithelial-specific markers. Immunohistochemical analyses showed that a putative stem cell marker, p63, a corneal epithelial differentiation maker, CK3, and a barrier function marker, Claudin-1, were expressed in the appropriate position in the cell sheets. These results suggest that the cell cartridge is effective for fabricating corneal epithelial cell sheets.
DOI	10.1002/term.1639
PMID	23239605