Satellite cell (SC) proliferation is observed following erythropoitin treatment in vitro in murine myoblasts and endurance training in vivo in human skeletal muscle. The present study aimed to investigate the effects of prolonged erythropoiesis-stimulating agent (ESA; darbepoetin-α) treatment and endurance training, separately and combined, on SC quantity and commitment in human skeletal muscle. Thirty-five healthy, untrained men were randomized into four groups: sedentary-placebo (SP, n = 9), sedentary-ESA (SE, n = 9), training-placebo (TP, n = 9) or training-ESA (TE, n = 8). ESA/placebo was injected once weekly and training consisted of ergometer cycling three times a week for 10 weeks. Prior to and following the intervention period, blood samples and muscle biopsies were obtained and maximal oxygen uptake (Symbol) was measured. Immunohistochemical analyses were used to quantify fibre type specific SCs (Pax7+), myonuclei and active SCs (Pax7+/MyoD+). ESA treatment led to elevated haematocrit, whereas endurance training increased Symbol. Endurance training led to an increase in SCs associated with type II fibres (P < 0.05), whereas type I fibres showed no changes. Both ESA treatment and endurance training increased Pax7+/MyoD+ cells, whereas only ESA treatment increased the total content of MyoD+ cells. Epo-R mRNA presence in adult SC was tested with real-time RT-PCR using fluorescence-activated cell sorting (CD56+/CD45−/CD31−) to isolate cells from a human rectus abdominis muscle and was found to be considerably higher than in whole muscle. In conclusion, endurance training and ESA treatment may separately stimulate SC commitment to the myogenic program. Furthermore, ESA-treatment may alter SC activity by direct interaction with the Epo-R expressed on SCs.