Establishing a reproducible method for the culture of primary equine corneal cells and evaluation of antifungal drug effects on equine corneal keratocytes

For culture of primary equine corneal epithelial cells, keratocytes and endothelial cells, corneas from horses recently euthanized for reasons unrelated to the study were collected and enzymatically separated into individual layers for cell capture. The cells were plated and grown in culture with complete media. The cells were passaged once confluent and continued for at least three passages. Each cell type was characterized and described with morphologic descriptions and immunocytochemical staining. All three equine corneal cell types were successfully grown in culture. Equine keratocytes and endothelial cells were able to be cryopreserved and recovered. Effects of different antifungal drugs and delivery vehicles on equine corneal keratocyte morphology and proliferation were evaluated. This work provides a construct for reproducible cell culture of all three equine corneal cells and is designed to describe cell collection, morphology, cytoskeletal expression and characteristics in culture.