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Andrew Tsin, Keli Wright, Brandi S Betts-Obregon; Glucose Effect on RhREC Viability and VEGF Secretion. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1534. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Previous studies have shown that, in diabetic patients, there is an increase of retinal capillaries associated with the development of diabetic retinopathy in the eye. We hypothesize that cultured Rhesus Monkey Retinal Endothelial Cells (RhREC’s) respond to high glucose with an increase in cell proliferation as well as an increase in VEGF secretion.
RhREC’s were seeded at 20,000 cells/ml and incubated with 0 mM (hypoglycemic), 5.5 mM (euglycemic), 18.5mM (hyperglycemic) and 30 mM (hyperglycemic) glucose for 0, 24, 48 and 72 hours. Viable cells were counted using trypan blue dye exclusion method. ELISA were used to measure VEGF concentrations in cell media.
Compared to the number of viable cells in euglycemic control (5.5mM), at 24 hours, viable cell numbers decreased by 45% at 0 mM, increased by 40% at 18.5 mM and decreased by 67% at 30 mM. At 48 hours, viable cell numbers decreased by 45% at 0 mM, decreased by 56% at 18.5 mM, and decreased by 78% at 30 mM. At 72 hours, viable cell numbers decreased by 95% at 0 mM, decreased by 41% at 18.5 mM and decreased by 93% at 30 mM. In contrast to the glucose effect on viable cell numbers, increase in glucose concentration resulted in an increase in VEGF levels (pg/mL) in the cell media
Our results show that hyperglycemic treatments result in a decrease in viable cell number. Also, cells cultured in hyperglycemia secreted more VEGF than those cultured in physiological conditions.
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