Glucose is important role for cellular functions. However, the behavior of osteoblasts in low glucose condition remains unclear. Therefore, this study aimed to elucidate the effects of low glucose conditions on the biological functions of osteoblasts. MC3T3-E1cells as one of osteoblast-like cells were cultured in MEM with five different glucose concentrations (0, 25, 50, 75, and 100 mg/dl). Then, low glucose conditions decreased the live cell number, proliferation, and migration of MC3T3-E1 cells. Alkaline phosphatase activity, mineralization, runx2, and osteocalcin production were also suppressed, whereas autophagy and extracellular signal-regulated kinase expression was increased under low glucose conditions. These results suggest that glucose may regulate osteoblast proliferation and differentiation.