A new isolation procedure for bovine retinal pigment epithelial cells has been developed. It is based on perfusion of the whole bovine eye via the central ophthalmic artery with a cold, buffered isotonic salt solution free of divalent cations for 15 min. The perfusion both weakens the association of the pigment epithelial cells with Bruch's membrane and the adhesion between retina and pigment epithelium. The retina then is removed carefully, after which the pigment epithelial cells are detached from the Bruch's membrane by gentle jets of buffer solution. The perfusion technique provides a high yield of intact retinal pigment epithelial cells, which show good viability in subsequent cell culture. Hence, cells isolated in this way are not only very well suited for long-term cell culture but also for direct biochemical analysis and short-term incubation studies.
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