Optimization of Storage Temperature for Cultured ARPE-19 Cells
Figure 7
Cultured RPE cells were stored at 12°C, 16°C, and 20°C for seven days, and expression of actin, ZO-1, RPE65, PCNA, and caspase-3 was assessed. The percentage of cells expressing RPE65, PCNA, and caspase-3 was quantified by two independent and blinded investigators. (a) Photomicrographs showing immunostaining with phalloidin-Alexa 568 used to visualize actin filaments (red). Nuclei were stained with DAPI (blue). Original magnification: ×200. (b) Photomicrographs showing immunostaining of ZO-1 (green). Nuclei were stained with DAPI (blue). Original magnification: ×200. (c) Photomicrographs showing immunostaining of RPE65 (red). Nuclei were stained with DAPI (blue). Original magnification: ×200. (d) Bar chart demonstrating RPE65 expression in stored and control cells. Expression of RPE65 was maintained after storage at all three temperatures. Error bars: standard deviation of mean values. (e) Photomicrographs showing immunostaining of PCNA (red) in control and stored cells. Nuclei were stained with DAPI (blue). Original magnification: ×200. (f) Bar chart displaying the percentage of PCNA+ cells in the control cultures and in the storage groups. PCNA expression was maintained at 12°C and 16°C and increased after storage at 20°C compared to the control. Error bars: standard deviation of mean values. (g) Photomicrographs of cells stained with anti-caspase-3 antibody (red). Nuclei were stained with DAPI (blue). Original magnification: ×200. (h) Bar chart showing the percentage of caspase-3+ cells. There was no increase in caspase-3+ cells after storage compared to control. Error bars: standard deviation of mean values.
