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Titel	FRO: Invitro culture charachteristics of corneal epithelial cells
Abstract Nr.	262

Doel	To characterize corneal epithelial cells that have been freshly isolated and to perform a comparative analysis after a period of invitro culture.
Methodes	Corneal epithelium was isolated using an overnight incubation with dispase at 4°C. The resulting epithelial sheets were dissected away from the underlying stroma under a dissecting microscope. The sheets were trypsinized to render single cell suspensions. Half the cells were stained with the following antibodies and analyzed using flow cytometry: CD14, CD15, CD133, CD29, CD73, CD166, CD227, CD107a, CD90, CD45,CD34 and ABCG2. The other half were placed in culture and then analyzed after a period of invitro cultivation.
Resultaten	The main population of freshly isolated cells stained CD166+/-, CD73-, CD90-, CD14- and CD45-. A subset of cells had the phenotype CD45+ CD14+ CD166+ and another subset was CD45- CD90+ CD73+. After a 2 week period of invitro culture the major population stained CD166+, CD73+/-, CD90-, CD14-, CD45-. The CD45+ CD14+ CD166+ subpopulation had disappeared but the CD45- CD90+ CD73+ subpopulation remained.
Conclusie	We can conclude that there are CD45+ CD14+ CD166+ haematopoietic cells present initially, but they are selectively removed after a 2 week invitro culture. However there is another CD45- CD90+ CD73+ subpopulation, possibly a mesenchymal one that remains a contaminant of the corneal epithelial cell culture.

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