Canine distemper virus (CDV), a highly contagious morbillivirus, continues to pose a major threat to domestic and wild carnivores worldwide. Efficient isolation of field strains is often limited by the lack of viral entry receptors in conventional cell culture systems such as Vero cells. To overcome this limitation, we established a recombinant Vero cell line stably expressing the canine signaling lymphocyte activation molecule (SLAM/CD150) receptor, the primary lymphoid entry receptor for CDV. The canine SLAM gene was amplified from peripheral blood mononuclear cells, cloned into the pTargeT mammalian expression vector, and transfected into Vero cells. SLAM expression was confirmed at the molecular level by PCR and further validated functionally using a Turkish field isolate (CDV-34388). Infected Vero-DogSLAM cells exhibited pronounced cytopathic effects (CPE), including extensive syncytium formation, while control Vero cells showed no evidence of CDV infection or detectable CPE. Importantly, immunofluorescence microscopy using anti-CDV polyclonal antibodies demonstrated specific intracellular viral antigen localization, providing protein-level confirmation of CDV entry and replication in SLAM-expressing cells. These findings demonstrate that the Vero-DogSLAM cell line markedly enhances the susceptibility of Vero cells to CDV infection, ensuring earlier detection of CPE and stable virus propagation for at least five passages. This recombinant system offers a robust platform for the isolation of field strains, diagnostic assay development, and potential vaccine research. Incorporation of immunofluorescence validation further strengthens its utility by providing visual evidence of virus–receptor interaction. In conclusion, engineering Vero cells to express canine SLAM significantly improves CDV isolation efficiency and provides an essential tool for advancing research in veterinary virology and immunodiagnostics.