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Both granulocyte colony-stimulating factor (G-CSF) and dexamethasone (DXM) are used for neutrophil (PMN) mobilization and collection. This prospective study was aimed to evaluate and compare the rate, severity and clinical significance of adverse reactions of these drugs alone and in combination in healthy donors. PMN mobilization was carried out using dexamethasone alone (8 mg orally; n=25) or glycosylated G-CSF alone (Lenograstim, 5 μg/kg subcutaneously, n=24) or in combination (n=23) prior to a standard granulocyte apheresis on the Spectra cell separator. The number of PMNs counted in the mobilized peripheral blood of the donors was 7.0 (3.6–20.4) ×109/L (DXM), 25.2 (15.5–49.7) ×109/L (G-CSF), and 31.6 (20.0–43.0) ×109/L (G-CSF+DXM), corresponding to PMN apheresis yields of 13 (8–43) ×109/U, 56 (34–118) ×109/U, and 83 (33–117) ×109/U, respectively. The three groups had comparable percentages of donors with at least one adverse effect (ranging from 75 to 80%), but the G-CSF-containing regimens were generally more toxic, as was reflected by higher percentages of donors with moderate to severe adverse reactions and higher overall severity scores of 2.28 (G-CSF) and 2.08 (G-CSF+DXM) compared with 1.33 in the DXM group (p≤0.001). With G-CSF alone, pain symptom complexes were more frequent, more severe, and more often triggered requests for analgesics (9/47 donors; 19%) and unwillingness to give further neutrophil donations (2/47 donors; 4%). The addition of DXM to G-CSF diminished some symptoms, particularly bone pain, headache and the frequency of requests for analgesics. The predominant symptoms in the DXM alone group were mild gastrointestinal complaints. We conclude that G-CSF stimulation improved neutrophil mobilization and apheresis yields at the expense of donor tolerability. Compared with G-CSF alone, the combination G-CSF and DXM did not increase the quantity or the severity of donor symptoms.