| Project/Area Number |
06454497
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | Ehime University School of Medicine |
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Principal Investigator |
OHASHI Yuichi Ehime University School of Medicine, Professor, 医学部, 教授 (00116005)
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| Co-Investigator(Kenkyū-buntansha) |
HANDA Hiroshi Tokyo College of Technology, Professor, 生命理工学部, 教授 (80107432)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1994: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Corneal epithelium / Immortalization / Corneal Transplantation / Three dimension culture / Functional differentiation / SV40 / SV 40-アデノウイルスベクター / 不死化細胞 / サイトケラチン / アルデヒドデヒドロゲナーゼ / 細胞培養 |
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| Research Abstract |
We have succeeded in immortalizing human corneal epithelial cells for the first time by using a recombinant SV40-adenovirus vector. These immortalized cells retained several properties consistent with epithelial cells, including formation of microvilli and desmosome as well as existence of aldehyde dehydrogenase. RT-PCR has demonstrated the expression of type 1 transglutaminase in these cells. Also, cornifin was found positive by immunohistochemical technique at the area where the cells became stratified. The expression of cadherin was increased in the presence of substance P,suggesting this cell line can be used for studying the machanism of adhesion and differentiation of corneal epithelium. When seeded onto a human keratocyte-containing type II collagen matrix, these immortalized corneal epithelial cells formed a stratified layr. This reconstructed tissue may be applicable as a transient substitute of corneal epithelial cells. In addition, in vitro test for screening the cytotoxicity of eyedrop solution has been developed in our laboratory using lactic dehydrogenase as a indicator, and has disclosed the cytotoxicity of unoprostone and timolol to corneal epithelial cells as has been experienced in clinical situation. This particular cell line has been widely distributed to number of research laboratories and extensively used as a source of human corneal epithelial cells.
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