Recombinant human erythropoietin (rHuEPO) has been used as a performance-enhancing agent by athletes in a variety of sports. The resulting increase in hematocrit levels leads to increased blood viscosity and can affect blood flow, potentially increasing the athlete's risk of developing health complications. However, the actual effects of using rHuEPO on microvascular blood flow and post-occlusive reactive hyperemia are currently unknown. We therefore evaluated the effect of rHuEPO on the cutaneous microcirculation in well-trained cyclists using laser speckle contrast imaging (LSCI). This study was part of a randomized, double-blind, placebo-controlled, parallel trial designed to investigate the effects of rHuEPO in 47 well-trained adult cyclists (age 18-50 years). Subjects received a weekly dose of either rHuEPO or placebo for 8 weeks, and LSCI was performed at baseline, after a maximal exercise test in week 6, and before maximal exercise in week 8. Endpoints included basal blood flux, maximum post-occlusion reperfusion, and time to return to baseline. Despite an increase in hematocrit levels in the rHuEPO-treated group, we found no statistically significant difference in microvascular function measured between the rHuEPO-treated group and the placebo group. Our results suggest that the increased hematocrit levels in rHuEPO-treated well-trained cyclists are not associated with changes in microvascular blood flow or post-occlusive reactive hyperemia measured using LSCI.