Some crude drugs have been known to useful effect of bone tissue regeneration in oriental medicine. These drugs were studied many scientific approach for bone formation. In this study, Alizarin Red S assay, and Alkaline phosphatase (ALP) activity of the seed of Hei Chou ethanol extract was investigated to confirm the osteoblastic effect. The result was observed that the seed ethanol extract enhances a significant induction of ALP activity, alizarin red S assay when differentiated osteogenic from MC3T3-E1 cell. In conclusion, Hei Chou extracts increased ALP activity, alizarin red S assay activity for bone formation. These results suggest that Hei Chou acts a successful drug for osteogenic differentiation in vitro and has the potential for use as a therapeutic agent for bone formation.