ISOLATION AND CULTURE OF IRIS PIGMENT-EPITHELIUM FROM IRIDECTOMY SPECIMENS OF EYES WITH AND WITHOUT EXFOLIATION SYNDROME

Objective: To culture iris pigment epithelium (IPE) from surgical irid ectomy specimens of eyes with and without exfoliation syndrome. Method s: The IPE was treated to obtain a single cell suspension. Cells were cultured in Ham F12 nutrient mixture, which was supplemented with 30% fetal bovine serum, 50-mg/mL gentamicin, and 2-mmol/L glutamine. After confluence, the cells were detached using a 0.125% trypsin-0.01% edet ic acid solution, resuspended, diluted, and subcultured. The IPE from primary cultures and subcultures was studied by transmission electron microscopy. Immunocytochemical staining was performed. Results: In the primary cultures of IPE from patients with exfoliation syndrome, curv ed, cross-banded, fine fibrils (diameter, 10-15 nm; periodicity, 10-14 nm) were found on the cell surface. Thicker fibrils (diameter, 24-48 nm; periodicity, 24-36 nm) were found external to the fine fibrils. Su bcultures contained mainly fine fibrils. The IPE cells stained positiv ely with anticytokeratin, S100 protein, and vimentin antibodies. Concl usion: Iris pigment epithelium can be successfully cultured from eyes with exfoliation syndrome. Studying the production of exfoliation mate rial in vitro should provide information about the pathogenesis of exf oliation syndrome and about the nature of the exfoliation material. Th e cultivation of normal IPE from surgical specimens provides a source for the study of the growth regulation and pharmacophysiology of IPE i n vitro.