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H. Kunikata, T. Abe, H. Murata, Y. Sagara, R. Wakusawa, H. Sato, M. Yoshida, N. Fuse, M. Tamai, K. Nishida; Hypothermia of 8 Degrees C Protects Cultured Retinal Pigment Epithelial Cells and Retinal Ganglion Cells Against Trypan Blue Toxicity . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2051.
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© ARVO (1962-2015); The Authors (2016-present)
To determine whether trypan blue (TB) is toxic to cultured human retinal pigment epithelial (ARPE–19) cells and rat retinal ganglion cells (RGC–5), and whether hypothermia of 8°C can protect these cells against the TB toxicity.
ARPE–19 cells and RGC–5 were cultured and exposed to balanced salt solution (BSS) as control, and to 0.05% and 0.5% TB in BSS at 37°C and 8°C for 1 minute. The percentage of ARPE–19 cells and RGC–5 that survived in the experimental groups relative to the control group was determined by resazurin 1 day after the exposure.
TB induced a statistically significant decrease in the percentage of RPE cell surviving at 0.5% TB at 37°C (P <0.01). The percentage of surviving ARPE–19 cells in 0.05% TB at both 37°C and 8°C, and in 0.5% TB at 8°C was not significantly different from that of controls. On the other hand, TB induced a statistically significant decrease in the percentage of RGC–5 surviving at all conditions except for 0.05% TB at 8°C (0.05% 37°C; P <0.05, 0.5% 37°C and 8°C; P <0.01).
These results indicate that TB is toxic to ARPE–19 cells and RGC–5 in culture, but reducing the temperature to 8°C has a protective effect against the TB toxicity.
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