Canine ocular papillomas occur on the haired skin of eyelids, conjunctival epithelium, and rarely on the cornea. Using PCR typing assays with canine papillomavirus type-specific primer sets, our study confirmed that the papillomas contained canine papillomavirus type 1. The positive result from a rolling circle amplification assay indicated the CPV1 viral genome in the cells is a circular episomal form. We also successfully established the first canine corneal cell line using the conditional reprogramming method. The cells exhibited an epithelial cell morphology, grew rapidly in vitro, and could be maintained long term. For the continued growth of the canine corneal cells, feeder cells played a more important role than Rho-kinase inhibitor Y-27632. More importantly, the viral CPV1 genome was maintained in the canine corneal cells during the long-term expansion. Unlimited supplies of these cells provide as a model for the study CPV in dog cells, and a platform for drug screening for effective therapies against canine papillomavirus infection in the future.