IMMORTALIZATION OF RABBIT CORNEAL EPITHELIAL-CELLS BY A RECOMBINANT SV40-ADENOVIRUS VECTOR

Purpose. Cultured corneal epithelial cell is detrimental because of it s short life span and its heterogeneity. We have tried to establish an immortalized epithelial cell line. Methods. Primary cultured rabbit c orneal epithelial cells were infected with a recombinant SV40-adenovir us vector and were cloned three times. Results. The immortalized cell continued to grow by more than 400 generations through 100 passages. S V40-associated large T antigen was demonstrable on the nuclear membran e of these immortalized cells by immunofluorescence technique. This ce ll line exhibited a similar cobblestone-like appearance as normal corn eal epithelial cells. Transmission electron microscopy showed a line o f evidence for stratification, including desmosome formation and micro villi development at the superficial cell layer. As the culture grew, these cells began to express cornea-specific 64 kD cytokeratins. In co ntrast to cultured normal corneal epithelial cells, this cell line had a good proliferative ability after a long-term storage in liquid nitr ogen. Conclusions. Because this particular cell line shares properties consistent with normal corneal epithelial cells and is easy to handle in vitro, it may serve as a useful tool in corneal epithelial researc h.