Establishment of a Canine Corneal Endothelial Cell Line Using Simian Virus 40 Induction

This study was performed to establish an immortalized canine corneal endothelial cell (CEC) line and characterize its properties to overcome the current limitations in sourcing primary CECs for basic and translational research in veterinary ophthalmology.

Eyeballs were obtained from dogs euthanized for reasons unrelated to this study. The canine corneas were dissected and isolated, and CECs were enzymatically dispersed and cultured. The cultured CECs were transfected with the simian virus 40 large T antigen gene for immortalization. Morphological evaluation of the immortalized cells and immunohistochemical analysis of CEC markers, including zonula occludens-1 and sodium-potassium adenosine triphosphatase, was conducted to confirm the identity of the cultured cells. Furthermore, post-passage morphological changes and cell proliferative ability were assessed.

Immortalized canine CECs displayed a small, cobblestone-like morphology. The cells proliferated in vitro and expressed zonula occludens-1 and sodium-potassium adenosine triphosphatase. Following repeated passaging, the cells maintained their morphological characteristics and proliferative ability.

Our immortalized canine CECs showed a high proliferative ability and characteristics similar to those of primary canine CECs, maintaining these properties for at least up to passages 25-30. This model may be valuable for studies aimed at optimizing culture conditions, testing therapeutic agents, and exploring cell-based therapies in veterinary ophthalmology.