Objective　To observe the effect of different concentrations of cytochalasin D (Cyto D) on the differentiation ability of human adipogenic stem cells to osteogenesis and adipogenesis.　Methods　Different concentrations of Cyto D that 0.05 μg/ml, 0.1μg/ml and 0.2μg/ml were added to growth medium, osteogenic differentiation medium and adipogenic differentiation medium, respectively, to disturb the elongation of cytoskeletal actin in different environments. Then the changes of cell differentiation ability were detected by oil red O staining, ALP staining and CCK8, in the treatment of 1d, 4d and 7d.　Results　Cyto D can both induce the formation of lipid droplets and change the basal state of osteogenesis in hASC. The combination of Cyto D and adipogenic differentiation medium can greatly promote the lipids formation. Low concentrations Cyto D increased the number of lipid droplets, and high concentrations Cyto D promoted the maturation of fat droplets. In combination with osteogenic differentiation medium, Cyto D was inversely proportional to the concentration and osteogenic effect.　Conclusion　Cyto D can enhance the effect of differentiation medium. Low concentration Cyto D mainly promotes osteogenesis and high concentration of Cyto D mainly promotes adipogenesis. Our exploration on the effect of myosin on the differentiation of adipogenic stem cells induced by Cyto D can provide important biological information for studying the differentiation mechanism of adipogenic stem cells.