Culture of Human Retinal Pigment Epithelium to Study UV Effects

Tratsk K., Ahmann P., Thanos S.,
Westfälische Wilhelms-Universität Münster, Klinik und Poliklinik für Augenheilkunde, Abt. für Experimentelle Ophthalmologie (Münster)

Objectives: Retinal pigment epithelium (RPE) is causally involved in the development and progression of the age related macular degeneration (AMD). The mechanisms of AMD-development remain largerly unknown.
Methods: We first established an RPE-culture method with embryonic cells of chicken. Thereafter, RPE-cells from „post-mortem“ human dissociated cultures were established according standardised protocols. The cells were cultured over several days to weeks. UV-irradiation (UV-A and UV-B) occured under sterile conditions with an 100 HBO/mercury bulb attached to the dissecting microscope „Leica MZ III“. The time-dependence of irradiation effects was analysed with morphometric, immunohistochemical and apoptosis-detecting techniques.
Results: „Post-mortem“ RPE cell cultures are routinely prepared, and the cells show tissue-specific morphologies in vitro. Immunohistochemistry to PEDF was positive. UV-irradiation but not bright microscope light results in morphometrically measurable alterations of cell shape and size. The cells show partially swellings but also shrinkage reminiscent of apoptotically advancing degeneration. These effects are dependent on duration of irradiation.
Conclusions: Cultures of RPE seem to be reliable and sensitive models to study cell-damage with pathogenetic relevance to AMD.

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