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UV irradiation causes multiple cellular changes in cultured human retinal pigment epithelium cells

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Abstract

Background

The retinal pigment epithelium maybe causally involved in the development and progression of age-related macula degeneration; however, the mechanisms leading to the development of age-related macula degeneration remain largely unknown. The purpose of this study was to examine cellular changes in the retinal pigment epithelium induced by direct irradiation with UV light in culture.

Methods

Retinal pigment epithelium cells from post-mortem human retinas were used to obtain dissociated cultures with cells retaining the ability to differentiate in vitro. These cells were cultured over several days to weeks. The UV radiation (UV-A and UV-B) occurred under sterile conditions with a 100 HBO/mercury bulb attached to a dissecting microscope, delivering co-axial illumination. The time dependence of irradiation effects was analysed using morphometric, immunohistochemical, functional and apoptosis-detecting techniques.

Results

Vital and proliferating retinal pigment epithelium cell cultures could be prepared consistently. The cells showed tissue-specific morphologies in vitro for several days to weeks. Pigment epithelium-derived factor was detected in these cells using immunocytochemistry and Western blots. The UV irradiation but not white light resulted in measurable alterations of cell shape and size. The irradiated cells showed partial swelling and shrinkage reminiscent of progressing apoptotic degeneration. TUNEL staining revealed that apoptosis was induced by UV light, but not detectably by white light. The phagocytosis of fluorescent micro-particles diminished after irradiation. These effects were dependent on the duration of irradiation.

Conclusions

Cultures of retinal pigment epithelium are suitable and sensitive models to study cell damage and may contribute to unravelling the pathogenetic mechanisms of retinal degeneration.

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Acknowledgements.The authors thank P. Ahmann for assistance with the in vitro experiments, L. Tatenhorst from the Institute of Neuropathology for providing the U-373 cells, and A. Rohlfing, Institute of Medical Physics and Biophysics, for measurements of the irradiation. I. Romann is acknowledged for assistance with the TUNEL staining. P. Vogt and C. Harmsen are acknowledged for providing the material from the Eye Bank of the University Eye Hospital Muenster. M. Pinheiro (University of Muenster, Germany) is acknowledged for typing the manuscript. K. Tratsk was a recipient of a CAPES fellowship from the Brazilian Government in collaboration with the DAAD. The work was supported by the IZKF Muenster (project F5 to S.T.)

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Authors and Affiliations

  1. University Hospital, University Federal of Santa Catarina, Campus Universitário, 88040-970, Florianópolis SC, Brazil

    Karla S. Tratsk

  2. Department of Experimental Ophthalmology, School of Medicine, University Eye Hospital Muenster, Domagkstrasse 15, 48149, Muenster, Germany

    Karla S. Tratsk & Solon Thanos

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  1. Karla S. Tratsk
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  2. Solon Thanos
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Correspondence to Solon Thanos.

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Tratsk, K.S., Thanos, S. UV irradiation causes multiple cellular changes in cultured human retinal pigment epithelium cells. Graefe's Arch Clin Exp Ophthalmol 241, 852–859 (2003). https://doi.org/10.1007/s00417-003-0747-0

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  • DOI: https://doi.org/10.1007/s00417-003-0747-0

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