2011 Fiscal Year Final Research Report
Assay for establishment of highly polarized retinal pigment epithelial cells
| Project/Area Number |
22791674
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | Kagoshima University |
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Principal Investigator |
SONODA Shozo 鹿児島大学, 医歯学総合研究科, 助教 (20325806)
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| Project Period (FY) |
2010 – 2011
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| Keywords | 細胞・組織 / polarized cell / 網膜色素上皮 / 加齢黄斑変性 |
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| Research Abstract |
Our group attempted to establish a reproducible method for culturing highly polarized RPE and tried to elucidate the pathophysiological mechanism of retinal disease using polarized culture RPE.
Highly polarized RPE cells were grown on Transwell^<TM> filters(12 mm, i. d). Primary cultures of RPE cells on T75 flask were seeded on fibronectin-coated TranswellTM. RPE cells were cultured in 1% FBS for more than 4 weeks. As in native tissue, porcine RPE cells formed a monolayer, were well pigmented, and were arranged in a regular hexagonal array. The integrity of RPE layer was confirmed by expression of TJ proteins ZO-1 and occluding and well developed microvilli, localization of pigment on the apical side, nuclei on basal side, and presence of tight-junctional complexes by electron microscopy. Well-differentiated RPE cells secreted VEGF-A preferentially to the basolateral side of the tissue. With respect to application of polarized culture RPE system, after stimulation with TNF-a, TER decreased about 15% of reduction in comparison to control at 24 hours, which was abolished by anti-TNF-a antibody. This reduction was inhibited by SB203580 inhibitor, but not by any other inhibitors.
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[Presentation] NSAIDs点眼による脈絡膜血管新生抑制2011
Author(s)
吉永就正, 有村昇, 大塚寛樹, 竹之内和則, 貞村ゆかり, 野間聖, 川原幸一, 園田祥三, 橋口照人, 丸山征郎, 坂本泰二
Organizer
第115回日本眼科学会総会
Place of Presentation
東京国際フォーラム(東京都)
Year and Date
2011-05-13
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