1Hillenkamp J., 1Zhang J. J., 2Cunningham J. R., 1Hussain A. A., 1Marshall J.

Depts. of Ophthalmology1 and Pharmacology2, The Rayne Institute, St. Thomas´ Hospital, London, UK

Aim: To characterise the transport properties of a novel in-vitro human tissue culture model of age-related macular degeneration (AMD) developed by Dr. Zhang.
Methods: Primary cultured human retinal pigment epithelial (RPE) cells were grown on samples of human Bruch`s membrane (BM) which had been incubated for 4 weeks in calcium-free Hank`s balanced salt solution (HBSS) in order to fully denude BM of any residual epithelial cells and choroidal elements. As a first step in the functional characterisation of the model we compared diffusion of taurine across BM and hydraulic conductivity of BM before and after incubation in HBSS using modified Ussing chambers.
Results: Freshly isolated RPE cells seeded onto denuded BM showed typical in-vivo functions, developed tight junctions and a degree of polarity. Donor age of BM tissue ranged from 42 to 94 years. Hydraulic conductivity of BM ranged between 0.88 and 4.6 m/sec/Pa. In initial experiments there was no statistically significant difference in taurine diffusion or hydraulic conductivity before and after incubation in HBSS.
Discussion: This preparation allows evaluation of the role of the RPE and BM in serial nutritional transport between the choroidal blood supply and the photoreceptors. Preliminary studies have shown that the functional properties of BM in the model do not differ from those of freshly obtained BM. These data indicate that measurement of transport across BM in the model reflects in-vivo transport. Studies on vectorial transport across the RPE and the RPE-BM-complex are currently being undertaken in conjunction with in-vitro ageing of the RPE.

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