Retinal pigment epithelium from human eyes was maintained in organ culture, and DNA synthesis was visualized by autoradiography of cultures exposed to labelled thymidine. In Ham's F10 medium, labelling of cells was observed after 7 as well as after 14 days in vitro. At both stages the labelled cells were characteristically located adjacent to areas of epithelial damage. Cells remote from such areas did not incorporate thymidine, This pattern was observed in epithelia removed from the posterior pole as well as from the periphery of the fundus. Several types of routine media and media with human as well as foetal bovine serum (FBS) were found to support DNA synthesis in the cultured epithelium, labelled cells were not observed in epithelium maintained in Ham's F10 without serum, or with low serum concentration. Our study demonstrates, that DNA synthesis does take place in organ cultured human retinal pigment epithelium when the appropriate culture conditions are used. Our findings indicate that induction of DNA synthesis in the cells is not promoted by an indiscriminate effect of the medium but related to the presence of epithelial damage.