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K Araki, Y Ohashi, T Sasabe, S Kinoshita, K Hayashi, X Z Yang, Y Hosaka, S Aizawa, H Handa; Immortalization of rabbit corneal epithelial cells by a recombinant SV40-adenovirus vector.. Invest. Ophthalmol. Vis. Sci. 1993;34(9):2665-2671.
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© ARVO (1962-2015); The Authors (2016-present)
PURPOSE: Cultured corneal epithelial cell is detrimental because of its short life span and its heterogeneity. We have tried to establish an immortalized epithelial cell line. METHODS: Primary cultured rabbit corneal epithelial cells were infected with a recombinant SV40-adenovirus vector and were cloned three times. RESULTS: The immortalized cell continued to grow by more than 400 generations through 100 passages. SV40-associated large T antigen was demonstrable on the nuclear membrane of these immortalized cells by immunofluorescence technique. This cell line exhibited a similar cobblestone-like appearance as normal corneal epithelial cells. Transmission electron microscopy showed a line of evidence for stratification, including desmosome formation and microvilli development at the superficial cell layer. As the culture grew, these cells began to express cornea-specific 64 kD cytokeratins. In contrast to cultured normal corneal epithelial cells, this cell line had a good proliferative ability after a long-term storage in liquid nitrogen. 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 research.
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