Expression of CD44 isoforms correlates with the metastatic potential of osteosarcoma. CD44 is a family of cell surface glycoproteins that not only function as receptors to hyaluronan and other extracellular matrix ligands but also is implicated in the invasiveness and metastasis of malignancies. The expression of CD44 and its variant isoforms was evaluated by immunohistochemistry in primary osteosarcoma, and the metastatic significance and prognostic relevance were assessed. Fifty formalin-fixed, paraffin-embedded conventional osteosarcoma samples were immunostained with monoclonal antibodies to CD44s, CD44v5, and CD44v6. When more than 50% of the tumor cells were stained, the result was regarded as ++ (overexpression). Staining patterns of CD44 mainly were membranous with a minor cytoplasmic pattern contribution. CD44s was overexpressed in 16% (eight of 50), CD44v6 in 20% (10 of 50), and CD44v5 in 46% (23 of 50) of the osteosarcomas. Of 26 patients with metastasis, CD44s was overexpressed in six (23%), CD44v6 in four (15%), and CD44v5 in 17 (65%). Overexpression of CD44v5 correlated significantly with metastasis. The survival rate of the patients in the CD44v5-positive (overexpressed) group was significantly lower than that of the patients in the CD44v5-negative group. Overexpression of CD44v5 may play a role in the metastatic behavior of osteosarcoma and may be a prognostic parameter of osteosarcoma.