Chronic Kidney Disease‐Mineral and Bone Disorder (CKD‐MBD) is characterized by an increased fracture risk. Bone marrow mesenchymal stromal cells (BMSCs) may be involved in the pathogenesis of bone disease and, in view of their promising potential applications in bone tissue engineering, the effect of uremia on BMSCs regenerative potential represents a central issue. The present study evaluated in vitro the effect of a serum pool from hemodialysis patients on BMSCs to observe its influence on osteogenic differentiation. Besides alterations in spatial organization and cytotoxicity along with hyperproliferation, gene expression analysis suggested an impairment in the osteogenic differentiation. More importantly, Receptor activator of nuclear factor kappa‐B ligand (RANKL) was upregulated with a mild reduction in osteoprotegerin levels. In summary, uremic environment seems to impair BMSCs osteogenic differentiation. Moreover BMSCs themselves may enhance osteoclastogenesis, feasibly contributing to the altered bone remodeling in CKD‐MBD patients. J. Cell. Physiol. 232: 2201–2209, 2017. © 2016 Wiley Periodicals, Inc. BMSCs were cultured in presence of a serum pool from hemodialysis patients (HUS) to study the effect of uremia on BMSC behavior. HUS induced both hyperproliferation and cytotoxicity, with an alteration in the spatial organization of cells. Moreover, osteogenic differentiation was impaired: a reduced mineralization and an altered pattern of gene expression were observed, in particular upregulation of RANKL and reduction in OPG levels.