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Dan-Ning Hu, MD; Steven A. McCormick, MD; Robert Ritch, MD

Arch Ophthalmol. 1997;115(1):89-94.

Abstract
Objective
To culture iris pigment epithelium (IPE) from surgical iridectomy specimens of eyes with and without exfoliation syndrome.

Methods
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% edetic 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, curved, 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. Subcultures contained mainly fine fibrils. The IPE cells stained positively with anticytokeratin, S100 protein, and vimentin antibodies.

Conclusion
Iris pigment epithelium can be successfully cultured from eyes with exfoliation syndrome. Studying the production of exfoliation material in vitro should provide information about the pathogenesis of exfoliation syndrome and about the nature of the exfoliation material. The cultivation of normal IPE from surgical specimens provides a source for the study of the growth regulation and pharmacophysiology of IPE in vitro.

Author Affiliations
From the Tissue Culture Center, the Department of Pathology and Laboratory Medicine (Drs Hu and McCormick) and the Department of Ophthalmology (Drs Hu and Ritch), the New York Eye and Ear Infirmary, New York; and the New York Medical College, Valhalla, NY (Drs Hu, McCormick, and Ritch). None of the authors has a financial interest in any of the material presented.

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