Hypothermia Protects Cultured Human Retinal Pigment Epithelial Cells against Trypan Blue Toxicity

Vol. 220, No. 2, 2006

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Original Paper

Hypothermia Protects Cultured Human Retinal Pigment Epithelial Cells against Trypan Blue Toxicity
Hiroshi Kunikata^a, Hiroshi Tomita^c, Hiromi Murata^a, Yoshiko Sagara^a, Hajime Sato^a, Yuko Wada^a, Nobuo Fuse^a, Yoichi Nakagawa^{a, d}, Toshiaki Abe^b, Makoto Tamai^c

^aDepartment of Ophthalmology and Visual Science, and
^bDivision of Clinical Cell Therapy, Tohoku University Graduate School of Medicine, and
^cDepartment of Biofunctional Science, Tohoku University Biomedical Engineering Research Organization, Sendai, and
^dDepartment of Ophthalmology, Ishinomaki Red Cross Hospital, Miyagi, Japan

Address of Corresponding Author

Ophthalmologica 2006;220:114-117 (DOI: 10.1159/000090576)

Key Words

Hypothermia, protecting effect
Trypan blue toxicity
Retinal pigment epithelial cells

Abstract

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°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°C. The percentage of cells surviving at 4°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°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°C had a protective effect against higher concentrations or longer exposure durations of TB compared to exposure at 37°C.

Author Contacts

Hiroshi Kunikata, MD
Department of Ophthalmology and Visual Science
Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku
Sendai 980-8574 (Japan)
Tel. +81 22 717 7294, Fax +81 22 717 7298, E-Mail kunikata@oph.med.tohoku.ac.jp

Article Information

Received: April 22, 2005
Accepted after revision: July 28, 2005
Number of Print Pages : 4
Number of Figures : 2, Number of Tables : 0, Number of References : 23

Medline Abstract (ID 16491034)

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