Mesenchymal stem cells (MSCs) are multipotent cells that have potential to differentiate into various phenotypes and appear useful for therapeutic applications in regenerative medicine. Mesenchymal Stem Cell Basal Medium (MSCBM; Cambrex) is a widespread and suitable medium used in MSC cultivation, but it is extremely difficult to use generally for clinical treatment because of its unclear traceability and cost. Assessment of cost-effectiveness is a critical issue for successful practical application; therefore, we have evaluated the effects of a generally used medium, Dulbecco's Modified Eagle's Medium (D-MEM) on the expansion of MSCs in comparison with MSCBM. To isolate human MSCs, bone marrow aspirates were taken and cultured in MSCBM or D-MEM. Proliferation assay indicated that MSCs isolated in both media showed a similar growth rate. When supplemented with osteo-inductive reagents, alkaline phosphatase activity was not significantly different between cells in D-MEM and MSCBM. Moreover, the cells expressed identical mesenchymal lineage markers, but not endothelial and hematopoietic lineage markers. Our findings suggest that cells obtained from bone marrow and cultured in D-MEM might possess proliferative capacity and the potential to differentiate into an osteogenic lineage. In conclusion, D-MEM might be a suitable basal medium for the cultivation of MSCs for clinical applications.