We have characterized 4 human squamous carcinoma cell lines established from the larynx and hypopharynx area. All the cell lines were cultured in RPMI-1640 medium. During the growth they showed monolayer adherence pattern in culture flask. They showed tonofilament on transmission electromicroscopy which is characteristic of squamous cell epithelium. DNA finger-printing using Hinf-l proved them to be originated from different beings. Flow cytometric analysis revealed them to show aneuploidy. Immunohistochemical staining for cytokeratin was done using CK1, CK8.13, CK19 and CAM5.2 antibody, and produced various patterns of positivity. All the cell lines showed varying degrees of tumorigenecity in athymic nude mice when injected subcutaneously, but only heterotransplanted SNU-1041 cell line showed continuous tumor growth. Histopathologic findings of the heterotransplanted tumors were identical to those of the original tumors of patients. This study suggests that establishment of many different squamous cell lines might bestow great capability in researches of the head and neck cancer.