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Establishing and functional testing of a canine corneal construct
Author(s) -
Werner Anke,
Braun Michael,
Reichl Stephan,
Kietzmann Manfred
Publication year - 2008
Publication title -
veterinary ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.594
H-Index - 50
eISSN - 1463-5224
pISSN - 1463-5216
DOI - 10.1111/j.1463-5224.2008.00646.x
Subject(s) - cornea , in vivo , dexamethasone , cell culture , in vitro , immunohistochemistry , cell , chemistry , pathology , biology , ophthalmology , medicine , endocrinology , biochemistry , genetics , microbiology and biotechnology
Purpose To provide a model to be used for in vitro studies on drug effects in dogs, this study was conducted to establish a protocol for the construction of a three‐dimensional corneal construct. Primary canine corneal cells and a rabbit corneal epithelial (RCE) cell line were used in comparison. Methods The corneal construct was assembled step by step in membrane inserts of a six‐well plate over a total of 5 weeks, including culture at the air–liquid interface to allow a differentiation of the epithelial cells. The constructs were studied histologically. Single cell cultures of canine corneal cells as well as RCE cells were stimulated with lipopolysaccharides (LPS) and sodium dodecyl sulfate (SDS). Treatment with different concentrations of dexamethasone was used to test its effects on the cellular prostaglandin E 2 (PGE 2 ) production. The same experiments were repeated with the corneal constructs and the reactions compared. Expression of the glucocorticoid receptor (GR) in the constructs was studied using immunohistochemistry. Results A protocol for the construction of a vital corneal construct was established and the morphological similarity to the canine cornea in vivo shown. The GR protein was detected in all three cell types of the constructs. Stimulation with LPS and SDS led only in the corneal constructs to a significantly increased PGE 2 production, which could be reduced by dexamethasone. Conclusions The corneal construct is an interesting system to test drug effects on corneal cells. It allows studies on a cornea‐like system including all three major cell types.