An SV40-immortalized human corneal epithelial cell line and its characterization.
OBJECTIVE
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.
Araki-Sasaki
K
K
Division of Ophthalmology, Kinki Central Hospital, Hyogo, Japan.
Ohashi
Y
Y
Sasabe
T
T
Hayashi
K
K
Watanabe
H
H
Tano
Y
Y
Handa
H
H
D000444:Aldehyde Dehydrogenase / Q000378:metabolism; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007633:Keratins / Q000378:metabolism; D008875:Middle Aged; D011994:Recombinant Proteins / Q000378:metabolism; D013539:Simian virus 40
D000952:Antigens, Polyomavirus Transforming; D011994:Recombinant Proteins; D007633:Keratins; D000444:Aldehyde Dehydrogenase