title: A human corneal equivalent constructed from SV40-immortalised corneal cell lines.
abstract: Within the last decade, extensive research in the field of tissue and organ engineering has focused on the development of in vitro models of the cornea. The use of organotypic, three-dimensional corneal equivalents has several advantages over simple monolayer cultures. The aim of this study was to develop a corneal equivalent model composed of the same cell types as in the natural human tissue, but by using immortalised cell lines to ensure reproducibility and to minimise product variation. We report our success in the establishment of an SV40-immortalised human corneal keratocyte cell line (designated HCK). A collagen matrix, built up with these cells, displayed the morphological characteristics of the human stromal tissue and served as a biomatrix for the immortalised human corneal epithelial and endothelial cells. Histological cross-sections of the whole-cornea equivalents resemble human corneas in tissue structure. This organotypic in vitro model may serve as a research tool for the ophthalmic science community, as well as a model system for testing for eye irritancy and drug efficacy.
authors.affiliation: Institute of Biophysics, University of Bremen, Otto Hahn Allee 1, 28359 Bremen, Germany. michaela@Kruppa-toelz.de
authors.forename: Michaela
authors.initials: M
authors.lastname: Zorn-Kruppa
authors.forename: Svitlana
authors.initials: S
authors.lastname: Tykhonova
authors.forename: Gazanfer
authors.initials: G
authors.lastname: Belge
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bednarz
authors.forename: Horst A
authors.initials: HA
authors.lastname: Diehl
authors.forename: Maria
authors.initials: M
authors.lastname: Engelke
mesh_terms: D000199:Actins / Q000235:genetics; D000826:Animal Testing Alternatives / Q000379:methods*; D000957:Antigens, Viral, Tumor / Q000235:genetics; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D003315:Cornea / Q000166:cytology* / Q000821:virology; D004728:Endothelium, Corneal / Q000166:cytology / Q000821:virology; D019573:Epithelium, Corneal / Q000166:cytology / Q000821:virology; D005455:Fluorescent Antibody Technique; D006651:Histocytochemistry; D006801:Humans; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
chemical_list: D000199:Actins; D000957:Antigens, Viral, Tumor