Related Experiment Video
Updated: Aug 11, 2026
Cryopreservation of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells at the Optimal Stage
Published on: November 3, 2023
Hypothermia protects cultured human retinal pigment epithelial cells against trypan blue toxicity
Hiroshi Kunikata1, Hiroshi Tomita, Hiromi Murata
1Department of Ophthalmology and Visual Science, Tohoku University Graduate School of Medicine, Sendai, Japan. kunikata@oph.med.tohoku.ac.jp
Purpose:
To determine whether trypan blue (TB) is toxic to cultured human retinal pigment epithelial (ARPE-19) cells, and whether hypothermia can protect ARPE-19 cells against TB toxicity.
Methods:
ARPE-19 cells were cultured and exposed to balanced salt solution as a control, while other cells were exposed to 0.05, 0.2, and 0.5% TB dye at 37 and 4 degrees C for 5 and 30 min. The percentage of ARPE-19 cells that survived was determined by resazurin 1 day after the exposure.
Results:
A statistically significant decrease in the percentage of ARPE-19 cells surviving was found after exposure to 0.2 and 0.5% TB at any temperature or for any exposure duration (p < 0.01). The percentage of RPE cells surviving at 0.05% was not significantly different from that of controls except for a 30-min exposure at 37 degrees C. The percentage of cells surviving at 4 degrees C for a 5-min exposure to 0.5% TB and a 30-min exposure to 0.2 and 0.5% was significantly higher than that at 37 degrees C under each condition (p < 0.01 for all).
Conclusions:
These results indicate that TB is toxic to human RPE cells, and the toxicity is dose- and exposure duration-dependent. Exposing the cells at 4 degrees C had a protective effect against higher concentrations or longer exposure durations of TB compared to exposure at 37 degrees C.
