Certain breeds are known to be overrepresented among mast cell tumor (MCT) patients, but other risk factors have not been evaluated. This study presents results from a case-control study of 252 dogs with grade 2 or grade 3 cutaneous MCT. Increased risk for MCT development was found in spayed females (adjusted odds ratio [OR], 4.11), boxers (adjusted OR, 6.09), Labrador retrievers (adjusted OR, 3.95), pugs (adjusted OR, 3.17), golden retrievers (adjusted OR, 2.12), the mastiff and terrier phylogenetic cluster (adjusted OR, 3.19), and breeds classified as large (adjusted OR, 2.10) or giant (adjusted OR, 5.44). Additional studies are needed to evaluate the role of these and other potential risk factors in MCT development.