Human retinal pigment epithelium in long term explant culture

Abstract

Explants of human retinal pigment epithelium were maintained in culture in various types of media, and examined by light and transmission electron microscopy. After one month in vitro, the central areas showed a monolayered configuration with distinct polarity and presence of ruthenium red stainable material on the apical surface. On the peripheral areas of Bruch's membrane, multilayered lesions were observed to develop and to extend from the monolayered epithelium and past the cut edge in Bruch's membrane. Cells in these lesions contained little melanin and generally lacked an apico-basal polarity. Ruthenium red staining revealed the presence of electron dense material on the apical surface of the lesions as well as in the extracellular space between cells in the various layers. Development of multilayered lesions with deposition of extracellular material are seen in various chorio-retinal disorders, including senile macular degenerations and also subsequent to laser and cryo-therapy. The findings in the present study point to the explant culture system as a valuable tool in the study of important aspects of chorio-retinal pathology.

Argon laser lesions in the human RPE in vitro

“…I n the present study, pigment cells outside the lesion area retained several of the in vivo characteristics. l'his is in concordance with a previous organ culture study where Ham's F10 medium was demonstrated to support maintenance of' the epithelium in long-term culture (Nicolaissen J r et al 1986). It further indicates that the alterations observed in the lesion area are not effectuated by a modulating activity of the medium per se.…”

“…This finding confirms the stable character of the monolayered organization of the pigment epithelium. It is retained in vitro under a multitude of culture conditions on plastics (Hayashi et al 1978;Flood et al 1980;Basu et al 1983), collagen (Hall & Quon 1981), on Bruch's membrane (Tso et al 1973;Nicolaissen J r et al 1986), and on vitreous gels (Forrester et al 1986).…”