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 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.


CONCLUSIONS
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.
Hu
D N
DN
Department of Pathology and Laboratory Medicine, New York Eye and Ear Infirmary, New York, USA.
McCormick
S A
SA
Ritch
R
R
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques; D002455:Cell Division; D002469:Cell Separation; D002470:Cell Survival; D002648:Child; D002675:Child, Preschool; D003470:Culture Media; D003598:Cytoskeletal Proteins / Q000378:metabolism; D017889:Exfoliation Syndrome / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D018463:Filtering Surgery; D005902:Glaucoma, Open-Angle / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D007498:Iris / Q000378:metabolism / Q000473:pathology* / Q000601:surgery / Q000648:ultrastructure; D008875:Middle Aged; D010857:Pigment Epithelium of Eye / Q000378:metabolism / Q000473:pathology* / Q000601:surgery / Q000648:ultrastructure; D009418:S100 Proteins / Q000378:metabolism
D003470:Culture Media; D003598:Cytoskeletal Proteins; D009418:S100 Proteins