Simple procedure to form an implantable, well-differentiated monolayer of cultured retinal pigment epithelial cells

2014 Fiscal Year Final Research Report

• Project/Area Number: 24791867
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Ophthalmology
• Research Institution: Nagoya City University
• Principal Investigator: KATO Aki 名古屋市立大学, 医学(系)研究科(研究院), 助教 (60405157)
• Project Period (FY): 2012-04-01 – 2015-03-31
• 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

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.

Free Research Field

網脈絡膜疾患