MONOLAYERED EXPLANTS IN THE STUDY OF RETINAL PIGMENT EPITHELIAL BEHAVIOR IN CULTURE

Abstract

A technique that permits removal of viable retinal pigment epithelial (RPE) explants of determined size from Bruch's membrane, and the transfer of such explants with maintained apico-basal polarity to cell culture dishes is presented. The RPE is a polarized tissue where the apical surface is involved in the interchange of material between the choroid and sensory retina and in phagocytosis of visual receptor outer segments. The maintenance of this polarity is of importance in studies aimed at elucidating these functions on pure RPE explants in early primary culture. No previous work has presented a method that permits this maintenance. The possibility of standardizing the size of these explants should facilitate quantitative studies on phagocytosis and uptake of markers and labelled compounds. The described dissection procedure is also currently used to separate the RPE as a pure cell population from surgically removed chorio-retinal biopsies for cell culture purpose.