Culture and characterisation of epithelial cells from human pterygia
- Nick Di Girolamoa,
- Nicodemus Tedlaa,
- Rakesh K Kumara,
- Peter McCluskeya,
- Andrew Lloyda,
- Minas T Coroneob,
- Denis Wakefielda
- aInflammation Research Unit, School of Pathology, University of New South Wales, Australia, bDepartment of Ophthalmology, Prince of Wales Hospital, Sydney, 2052, Australia
- Dr Nick Di Girolamo, Inflammation Research Unit, School of Pathology, The University of New South Wales, Sydney, 2052, Australia.
- Accepted 18 May 1999
Abstract
BACKGROUND/AIMS Pterygia are a common disorder of the ocular surface. The disease represents a chronic fibrovascular and degenerative process thought to originate at the conjunctival-corneal junction, where altered limbal stem cells are proposed to be the cell of origin. Extensive epidemiological evidence exists to implicate ultraviolet B irradiation in the pathogenesis of pterygia. To date no animal or in vitro culture model has been developed to test such an hypothesis. The aim of this study was to establish and characterise a pure population of epithelial cells derived from pterygium tissue.
METHODS Tissue specimens were obtained from patients undergoing pterygium excision. Explants were cultured in either serum free or serum supplemented medium. Primary and passaged cells were processed for light microscopy, analysed by flow cytometry, and characterised immunohistochemically using specific antibodies.
RESULTS In serum free culture, cuboidal cells with typical morphology of epithelial cells migrated from the pterygium explants from 3 days onwards and eventually formed a cohesive monolayer. Passaged cells consisted of 98.4% cytokeratin positive cells and demonstrated immunoreactivity for multiple cytokeratins, including AE1, AE3, AE5, but were negative for AE8. These cells also expressed an epithelial specific antigen, together with vimentin and mucin, as did epithelial cells in sections of pterygia.
CONCLUSIONS A relatively simple method of isolating pterygium epithelial cells has been established. Cultured pterygium epithelial cells are phenotypically and functionally similar to their in vivo counterparts with respect to keratin, vimentin, and mucin expression. In vitro assays using these cells may aid in elucidating the pathogenesis of pterygia.
