Culture of feline corneal epithelial cells and infection with feline herpesvirus-1 as an investigative tool

Lynne S. Sandmeyer Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON, N1G 2W1, Canada.
Present address is Department of Small Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5B4, Canada.

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Charlotte B. Keller Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON, N1G 2W1, Canada.

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Dorothee Bienzle Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON, N1G 2W1, Canada.

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Abstract

Objective—To isolate and characterize pure cultures of feline corneal epithelial cells and to assess the extent and nature of feline herpesvirus (FHV)-1 infection in these cells.

Sample Population—Healthy eyes from 23 recently euthanatized cats.

Procedure—Stroma and epithelium of the rostral portion of the cornea were surgically isolated, and epithelial cells were detached from the stroma by enzymatic incubation. Epithelial cells were cultured in hormone-supplemented media. Cells were passaged, and cytokeratin expression was assessed. Cells were then infected with FHV-1, and cytopathic effects were determined.

Results—Cell cultures were readily established from samples obtained from each eye and could be maintained through 6 passages. Cultured cells expressed cytokeratins 3 and 12 but not other cytokeratins. Infection with FHV-1 was rapid and caused widespread cytopathic effects.

Conclusions and Clinical Relevance—Feline corneal cells cultured in vitro during multiple passages maintain consistent morphologic characteristics and intermediate filament expression. They are susceptible to infection with FHV-1 and may provide a useful in vitro model for investigation of ocular drugs. (Am J Vet Res 2005;66:205–209)

Abstract

Objective—To isolate and characterize pure cultures of feline corneal epithelial cells and to assess the extent and nature of feline herpesvirus (FHV)-1 infection in these cells.

Sample Population—Healthy eyes from 23 recently euthanatized cats.

Procedure—Stroma and epithelium of the rostral portion of the cornea were surgically isolated, and epithelial cells were detached from the stroma by enzymatic incubation. Epithelial cells were cultured in hormone-supplemented media. Cells were passaged, and cytokeratin expression was assessed. Cells were then infected with FHV-1, and cytopathic effects were determined.

Results—Cell cultures were readily established from samples obtained from each eye and could be maintained through 6 passages. Cultured cells expressed cytokeratins 3 and 12 but not other cytokeratins. Infection with FHV-1 was rapid and caused widespread cytopathic effects.

Conclusions and Clinical Relevance—Feline corneal cells cultured in vitro during multiple passages maintain consistent morphologic characteristics and intermediate filament expression. They are susceptible to infection with FHV-1 and may provide a useful in vitro model for investigation of ocular drugs. (Am J Vet Res 2005;66:205–209)

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