March 1995
Volume 36, Issue 3
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Articles  |   March 1995
An SV40-immortalized human corneal epithelial cell line and its characterization.
Author Affiliations
  • K Araki-Sasaki
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • Y Ohashi
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • T Sasabe
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • K Hayashi
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • H Watanabe
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • Y Tano
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
  • H Handa
    Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
Investigative Ophthalmology & Visual Science March 1995, Vol.36, 614-621. doi:
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    • Get Citation

      K Araki-Sasaki, Y Ohashi, T Sasabe, K Hayashi, H Watanabe, Y Tano, H Handa; An SV40-immortalized human corneal epithelial cell line and its characterization.. Invest. Ophthalmol. Vis. Sci. 1995;36(3):614-621.

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      © 2016 Association for Research in Vision and Ophthalmology.

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Abstract

PURPOSE: The authors attempted to immortalize human corneal epithelial cells; it is difficult to propagate primary human corneal epithelial cells because of scarcity of available tissue. However, cell immortalization by virus is always accompanied by shedding of free virus. The current study was performed to establish a cell line that produces no free viral particle. METHODS: Primary cultured human corneal epithelial cells were infected with a recombinant sv40-adenovirus vector and were cloned three times to obtain a continuously growing cell line. Morphologic, cytologic, and biochemical characteristics of this cell line were analyzed. RESULTS: This cell line continued to grow for more than 400 generations, exhibiting a cobblestone-like appearance similar to normal corneal epithelial cells in culture. Transmission electron microscopy showed the evidence for the characteristic features of epithelial cells, including desmosome formation and development of microvilli. It expressed cornea-specific, 64-kD cytokeratin in addition to five major insoluble proteins. By enzymatic analysis using NADP as a coenzyme and a gas chromatograph mass spectrometer, this cell line was found to possess 8.71 IU/mg protein of aldehydedehydrogenase activity. When this cell line was grown at air-liquid interface on collagen type I gel, it differentiated in a multilayered fashion. CONCLUSIONS: The authors have established an SV40-immortalized human corneal epithelial cell line with properties similar to normal corneal epithelial cells.

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