Human c‐mpl ligand or thrombopoietin (TPO) has been proved to be a critical cytokine in the physiological regulation of thrombopoiesis. Previous evidence suggested that TPO production is constitutive and TPO plasma levels are regulated by the platelet–megakaryocyte mass through c‐mpl receptor‐mediated uptake and metabolism. To evaluate whether this mechanism of TPO level regulation is also operative in subjects with an elevated platelet count, we evaluated serum TPO in 32 patients with thrombocytosis due to essential thrombocythaemia (ET) or polycythaemia vera (PV) and in 70 subjects with reactive thrombocytosis; 32 healthy subjects were also studied. TPO levels were significantly higher in the ET and PV groups (median 246.2 pg/ml, range 93.5–4596) and reactive thrombocytosis patients (median 287 pg/ml, range 82.7–1960.0) than in normal subjects (median 156.7 pg/ml, range 62.2–352.7). No significant difference was found between the two groups of patients, indicating that serum TPO levels cannot differentiate between primary and reactive thrombocytosis. No significant correlation was found between platelet count and TPO levels in either ET‐PV or reactive thrombocytosis, whereas a trend toward a correlation between acute‐phase reactants and TPO was observed in patients with reactive thrombocytosis. These results indicate that TPO clearance by platelets–megakaryocytes is not fully operative or is not the only mechanism of TPO level regulation in primitive and reactive thrombocytosis.