title: The immunohistochemical characterisation of an SV40-immortalised human corneal epithelial cell line.
abstract: Alternatives to the Draize rabbit eye irritation test are currently being investigated. Because of morphological and biochemical differences between the rabbit and the human eye, continuous human cell lines have been proposed for use in ocular toxicology studies. Single cell-type monolayer cultures in culture medium have been used extensively in ocular toxicology. In the present study, an SV40-immortalised human corneal epithelial (HCE) cell line was characterised immunohistochemically, by using 13 different monoclonal antibodies to cytokeratins (CKs), ranging from CK3 to CK20. The results from the monolayer HCE cell cultures were compared with those from the corneal epithelium of human corneal cryostat sections. Previous studies have shown that the morphology of the HCE cell is similar to that of primary cultured human corneal epithelial cells, and that the cells express the cornea-specific CK3. In the study reported here, we show that the cell line also expresses CKs 7, 8, 18 and 19. These CKs are typically expressed by simple epithelial cells, and are not found in the human cornea in vivo. Therefore, the monolayer HCE cell line grown in culture medium does not express the CK pattern that is typical of human corneal epithelium. This should be taken into consideration when using HCE cell cultures in similar single cell-type experiments for ocular toxicology.
authors.affiliation: Medical School, University of Tampere, 33014 Tampere, Finland.
authors.forename: Anne
authors.initials: A
authors.lastname: Huhtala
authors.forename: Sami K
authors.initials: SK
authors.lastname: Nurmi
authors.forename: Hanna
authors.initials: H
authors.lastname: Tähti
authors.forename: Lotta
authors.initials: L
authors.lastname: Salminen
authors.forename: Päivi
authors.initials: P
authors.lastname: Alajuuma
authors.forename: Immo
authors.initials: I
authors.lastname: Rantala
authors.forename: Heikki
authors.initials: H
authors.lastname: Helin
authors.forename: Hannu
authors.initials: H
authors.lastname: Uusitalo
mesh_terms: D023401:Animal Use Alternatives / Q000379:methods*; D000911:Antibodies, Monoclonal; D018929:Cell Culture Techniques; D002460:Cell Line; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000276:immunology / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D007633:Keratins