We have characterized various biologic, immunologic and growth properties of several cell lines established from a spontaneous rat sarcoma that was discovered more than 60 yr ago. The tumors consisted of mixed cell types with no detectable host cellular immune response. Cultures derived from single-cell clones of the parental cell line were non-invasive but highly tumorigenic even in adult rats. The cultured cells spontaneously released replication-competent endogenous rat type C virus which did not carry a transforming gene in its genome. Since normal cells from the same rat strain did not produce a retrovirus, it is possible that production of the endogenous retrovirus may have triggered specific cellular changes necessary for the oncogene expression and development of this tumor.