The effects of TSH on peroxidase activity (PO) and thyroglobulin (TG) production were investigated using primary cultures of thyroid cells obtained from patients with Basedow's disease (Basedow's cells). PO activity of cultured cells and TG concentration in the culture medium were measured by biochemical and sandwich enzyme immunoassays, respectively. The addition of TSH (10 mU/ml/day) to the medium did not increase the cell number but did increase the PO activity and TG concentration. It took more than 3 days for the PO activity of cells cultured with TSH (stimulated group) to reach a level twice that of cells cultured without TSH (control group), whereas 2 days of incubation with TSH was sufficient for increasing the TG concentration. When actinomycin D (AD) was added to the medium on the first day of 3-day incubation with TSH, the stimulatory effect of TSH on PO was completely blocked and the TG concentration was reduced to half that of the control group. AD given to the stimulated group on the last day of induction produced no inhibitory effect on the induction of PO activity by TSH, but reduced the TG concentration to almost half that in the stimulated group. An electron microscopic study of Basedow's cells cultured with AD and TSH failed to reveal any cytopathic change. The findings of the present study suggested that in cultured Basedow's cells TSH induces PO activity and TG production through the synthesis of new messenger RNA.