NDLI: A novel method of culturing human oral mucosal epithelial cell sheet using post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction.

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A novel method of culturing human oral mucosal epithelial cell sheet using post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction.

Content Provider	Semantic Scholar
Author	Oie, Yoshinori Hayashi, Ryuhei Takagi, Ryo Yamato, Masayuki Takayanagi, Hiroshi Tano, Yasuo Nishida, Kohji
Copyright Year	2010
Abstract	BACKGROUND/AIMS To cultivate human oral mucosal epithelial cell sheets with post-mitotic human dermal fibroblast feeder cells and modified keratinocyte culture medium for ocular surface reconstruction. METHODS Human oral mucosal epithelial cells obtained from three healthy volunteers were cultured with x-ray-treated dermal fibroblasts (fibroblast group) and NIH/3T3 feeder layers (3T3 group) on temperature-responsive culture dishes. Media were supplemented using clinically approved products. Colony-forming efficiency was determined in both groups. Histological and immunohistochemical analyses were performed for cell sheets. Cell viability and purity of cell sheets were evaluated by flow cytometry. RESULTS Colony-forming efficiency in the fibroblast group was similar to that in the 3T3 group. All cell sheets were well stratified and harvested successfully. The expression patterns of keratin 1, 3/76, 4, 10, 12, 13, 15, ZO-1 and MUC16 were equivalent in both groups. The percentage of p63-positive cells in the fibroblast group (46.1+/-4.2%) was significantly higher than that in the 3T3 group (30.7+/-7.6%) (p=0.038, t test). The cell viability and purity were similar between the two groups. CONCLUSION This novel culture method using dermal fibroblasts and pharmaceutical agents provides a safe cell processing system without xenogenic feeder cells for ocular surface reconstruction.
File Format	PDF HTM / HTML
DOI	10.1136/bjo.2009.175042
PubMed reference number	20538654
Journal	Medline
Volume Number	94
Issue Number	9
Alternate Webpage(s)	https://ir.library.osaka-u.ac.jp/repo/ouka/all/58220/24115_%E8%A6%81%E6%97%A8.pdf
Alternate Webpage(s)	https://doi.org/10.1136/bjo.2009.175042
Journal	The British journal of ophthalmology
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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