Isolation and culture of iris pigment epithelium from iridectomy specimens of eyes with and without exfoliation syndrome
D. N. Hu, S. A. McCormick and R. Ritch
Department of Pathology and Laboratory Medicine, New York Eye and Ear Infirmary, New York, USA.
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-micrograms/mL [corrected] 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.
