Purpose Corneal epithelial wound healing requires orchestrated cell proliferation, migration and adhesion. Disturbances in any or all of these events may underlie impaired corneal wound healing in diabetes. To separate these components, we have begun to characterize the effects of elevated extracellular glucose concentration on SV40 transformed human corneal epithelial cells (Araki-Sasaki, et ai IOVS, 1995, 36 614-621) Methods Cells were maintained in DMEM/F12 (1 1) or in this media containing 38mM glucose Adhesion Cells were allowed to adhere for 1 hour at 37°C to 96 well plates coated with extracellular matrix proteins. Non-adherent cells were removed by washing, then attached cells quantitated by using the fluorescent dye Calcein AM Migration: Cell migration in response to 10% fetal bovine serum was determined in blind well migration chambers. Results Adhesion Collagen 1 was the preferred substrate, then fibronectin = vitronectin > laminin (ECso 0 15+/-004, 3 95+/-1 9, 6 1+/-2 1 and >100ug/ml respectively, n=2or3). Pre-incubation of the cells with a monoclonal antibody to βl-integrin inhibited adhesion, indicating involvement of this protein in adhesion. Western blotting with an anti-βl polyclonal antibody confirmed the presence of this integrin in these cells. Exposure of the cells to 38mM glucose for i-5 weeks had no significant effect on adhesion. In contrast, cell migration was attenuated by up to 50% in cells that had been maintained in 38mM glucose Conclusions Supplementation of the growth media with glucose did not affect the adherent capabilities of SV40 transformed human corneal epithelial cells, but did lead to attenuated cell migration.
|Original language||English (US)|
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - 1997|
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