Br J Ophthalmol 1999;83:1077-1082
( September )
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
a Inflammation
Research Unit, School of Pathology, University of New South Wales,
Australia, b Department of Ophthalmology, Prince of Wales
Hospital, Sydney, 2052, Australia
Correspondence to: Dr Nick Di Girolamo, Inflammation Research Unit, School of Pathology,
The University of New South Wales, Sydney, 2052, Australia.
Accepted for publication 18 May 1999
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.
METHODSTissue
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.
RESULTSIn 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.
CONCLUSIONSA
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.
© 1999 by British Journal of Ophthalmology
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