Purpose: To isolate and characterize an epithelial cell (EC) line from a human donor cornea, which may serve as a reliable test cell line to address biomolecular issues and study the response of corneal epithelium to stressing events and therapeutic treatments.
Methods: A corneal button from a donor patient was treated with enzymes to separate the epithelial sheet and to free EC, which were put in tissue culture. ECs were characterized by optic and electronic microscopies, cytokeratins and PAX6 were detected by SDS-PAGE and western immunoblotting, the barrier function was evaluated by transepithelial electric resistance and by the immune detection of membrane junction proteins, and the karyotype was characterized according to the classical methods.
Results: Morphological analyses returned the picture of classical homogeneous polygonal morphology as expecetd by EC that was maintained over time and several in vitro passages. Transepithelial electric resistance values were characteristic of a typical barrier-forming cell line. The cytokeratin expression pattern was the one expected for corneal EC with a predominance of CK3 and CK5 and different from a human keratocyte cell line. The male karyotype showed 2 trisomies, of chromosomes 8 and 11.
Conclusions: All the data so far obtained with the HCE-F cells concur to certify this cell line as a stable, true primary human corneal EC line, which could then be used as a test cell line to study and address the questions concerning the biological response of human corneal epithelium to stressing and/or therapeutic treatments and as a term of comparison for EC derived from pathological corneas.