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We isolated human mesenchymal stem cells (hMSC) from dental pulp. These cells are multipotent and serve as precursors for various mesoderm-type cells such as osteoblasts, chondroblasts and adipocytes, as well as several other types of cells. This study investigated the possible role of gangliosides in osteoblast differentiation of hMSCs. Gangliosides, which exist in glycosphingolipid-enriched domains on the cell membrane The role of gangliosides that play in osteoblastogenesis is not yet clearly understood. Therefore, in this study, We investigated the relationship between gangliosides and osteoblast differentiation of human mesenchymal stem cells (hMSCs). The results of high-performance thin-layer chromatography (HPTLC) showed that ganglioside GD1a expression was increased during the differentiation of hMSCs into osteoblasts. Furthermore, we showed that involvement of alkaline phosphate (ALP) with GD1a by immunostaining. Taken together, these results suggest that gangliosides GD1a may play a role in the osteoblast differentiation process of hMSCs.