Podoplanin has been shown to be expressed in primary germ cell tumors (GCTs), with conflicting results regarding its specificity. However, podoplanin expression in metastatic GCTs and other metastatic tumors has not been extensively examined. The goal of this study was to evaluate the distribution and specificity of podoplanin using monoclonal antibody D2-40 in primary testicular and metastatic GCTs in comparison with other metastatic neoplasms. In total, 122 tumors were studied: 43 primary GCTs, 33 metastatic GCTs, 11 metastatic melanomas, 25 metastatic carcinomas, and 10 lymphomas. All foci of seminoma showed strong, diffuse membranous staining in more than 90% of cells in primary and metastatic GCTs. In contrast, other GCT components showed only focal cytoplasmic and/or partial membranous staining in a subset of cases. Among non-GCTs, only 1 metastatic melanoma, 1 lymphoma, and 3 metastatic carcinomas showed focal, weak cytoplasmic staining. Diffuse membranous immunoreactivity for podoplanin as detected by monoclonal antibody D2-40 is highly sensitive and specific for primary and metastatic seminoma. Immunodetection of podoplanin may be useful to support seminoma in the differential diagnosis of poorly differentiated epithelioid malignant neoplasms.