Simple procedure to form an implantable, well-differentiated monolayer of cultured retinal pigment epithelial cells
| Project/Area Number |
24791867
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Ophthalmology
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| Research Institution | Nagoya City University |
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Principal Investigator |
KATO Aki 名古屋市立大学, 医学(系)研究科(研究院), 助教 (60405157)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 網膜色素上皮単層シート / ブルッフ膜 / タイトジャンクション / 網膜色素上皮 / 加齢黄斑変性 / 移植 / シート / 再分化 / 黄斑部萎縮 / 単層シート / 免疫染色 / 電子顕微鏡 / ウェスタンブロット |
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| Outline of Final Research Achievements |
Light microscopy revealed the hexagonal shape of RPE cells with an overlying fibrous membrane, suggesting upside-down polarity of RPE and the sheet was easily detached from the bottom of the plate by the gentle pipetting technique. The RPE sheet expressed occludin and ZO-1 as tight junction markers as well as actin, collagen type IV and elastin as a Bruch’s membrane component in Western blotting. In SEM images, the upper side showed microvilli and lamellipodia of RPE cells and, thereafter, Bruch’s membrane-like structures involving elastic fibers and collagen. TEM disclosed the tight junctions of the RPE cells. Confocal immunofluorescence microscopy showed the expression of ZO-1 and occludin among RPE cells. The RPE sheet manufactured by our original method was well-structured as an implantable sheet with Bruch’s membrane and tight junction. This RPE sheet may have a potential as a better option for RPE transplantation.
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Report
(4 results)
Research Products
(2 results)
