Nonadherent Fcγ receptors (FcγR) expressing cells from SEYF-a tumors that form rosettes with sheep erythrocytes coated with IgG antibodies (EA) were isolated by Percoll density gradients. The EA-IgG rosette-forming cells were characterized by the parameters of 1) binding of IgG immune complexes; 2) binding of purified monoclonal antibodies against mouse FcR; 3) sensitivity to complement-dependent lysis mediated by syngeneic anti-SEYF-a antibodies; 4) expression of parental H-2 antigen when grown in F1 hybrids; 5) incorporation of [125I]5-iodo-2'-deoxyuridine; and 6) growth in syngeneic mice. The nonadherent EA-IgG rosette-forming cell population was found to be composed of both host lymhpocytes as well as of tumor cells. Tumor-seeking lymphocytes then were removed from SEYF-a tumors by velocity sedimentation on Percoll. The remaining cell population was tumorigenic and expressed FcR, as well as tumor antigens. These tumor EA-IgG rosette-forming cells exhibited a very low rate of DNA synthesis compared with that of nonrosetting tumor cells.