Herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) are members of the Herpesviridae family. About 40% to 80% of the world populations are infected with HSV and its prevalence is high in Iran. The high prevalence of this virus in the community and the ability of the virus in causing fatal diseases among immunocompromised patients, have encouraged studies to be performed on HSV and suitable cell lines which supports the propagation of HSV. The aim of this study was to evaluate the suitability of McCoy cell line in the isolation and propagation of HSV. An isolated wild-type HSV-1 was obtained from the labial vesicles of a 29-year-old patient who was referred to Ghaem Hospital (Mashhad, Iran). The virus was inoculated in McCoy cell monolayer cells and its titer was calculated by 50% tissue culture infectious dose (TCID50) method. Cytopathic effects (CPE) of HSV on McCoy cells appeared about 20 hours after the infection of cells. Titer of the virus was 10-5.25 TCID50/ml. Our data showed that the McCoy cell line supported the propagation of HSV in high titer. This was the first study that used McCoy cell line for the isolation and propagation of HSV-1. McCoy cell line could be used, as a proper cell line of HSV, for various studies in the future.