| Project/Area Number |
04454442
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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 | Kyoto Prefectural University of Medicine |
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Principal Investigator |
KINOSHITA Shigeru Kyoto Pref Univ Med, Ophth, Prof, 医学部, 教授 (30116024)
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| Co-Investigator(Kenkyū-buntansha) |
OHASHI Yuichi Ehime University, Ophth, Prof, 医学部, 教授 (00116005)
NISHIDA Kohji Kyoto Pref Univ Med, Ophth, Assist, 医学部, 助手 (40244610)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1992: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Corneal epithelium / Cell culture / Immortalization / Conjunctival epithelium / Epithelial transplantation / KGF / TGF-beta / 増植制御因子 / 株化 |
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| Research Abstract |
We aimed at developing effective treatment for primary ocular surface disorders. For this, in this project, we mainly focused on the methods controlling proliferation and differentiation of corneal epithelial cells. First, we established a noble cell culture system for freeze and long-term subculture of rabbit corneal epithelial cells. This culture media is composed of very low Ca^<2+> concentration with high aminoacid contents. Second, we established an immortalized rabbit corneal epithelial cell line. For this, primary cultured rabbit corneal epithelial cells were infected with a recombinant SV40-adenovirus vector and were cloned. This cell line shares properties consistent with normal corneal epithelial cells. Although these cells were not transferable as a epithelial transplantation at this point, our success has made a marked progress in handling corneal epithelial cells in vitro. Third, we investigated biological roles of several cell growth and/or regulatory factors. For instance, epidermal growth factor, which mainly comes from lacrimal gland, promoted corneal epithelial cell growth. Similar phenomenon was found in keratinocyte growth factor, but this was produced only in the corneal stromal cells. On the other hand, transforming growth factor -beta did inhibit corneal epithelial cell growth. We also studied spartial distribution of three TGF-beta isoforms in the cornea, and found TGF- beta 2 predominantly.
In the study of epithelial allograft rejection in vivo, we are now investigating IL-2 levels in tears in patients with cyclosporin treatment after corneal epithelial transplantation.
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