This work aimed to investigate the effect of Sophora flavescens polysaccharide on proliferation and osteogenic differentiation of dental pulp stem cells. The dental pulp stem cells were isolated from rat dental crown tissue and the surface biomarkers were identified by flow cytometry. The osteogenic differentiation of dental pulp stem cells was assessed by alizarin red staining and alkaline phosphatase staining. The dental pulp stem cells were treated with different doses of Sophora flavescens polysaccharide for different time points and cell viability was assessed by cell counting kit 8. Cell migration was detected by Transwell and wound healing assay. The expression of collagen I, osteocalcin I, runt-related transcription factor 2 and beta-catenin were measured by quantitative real time polymerase chain reaction assay and Western blotting assay. The cluster of differentiation 90 was highly expressed and cluster of differentiation 45 expressions was relatively low on surface of isolated dental pulp stem cells. Sophora flavescens polysaccharide suppressed the proliferation of dental pulp stem cells. The treatment with low dose Sophora flavescens polysaccharide (0.2 mg/ml) promoted the migration and osteogenic differentiation of dental pulp stem cells, but higher concentration of Sophora flavescens polysaccharide (1 and 10 mg/ml) suppressed the migration and osteogenic differentiation of dental pulp stem cells. Treatment with low concentration of Sophora flavescens polysaccharide up-regulated the expression of osteogenic biomarkers. Low dose Sophora flavescens polysaccharide can promote the migration and osteogenic differentiation of dental pulp stem cells and up-regulated the expression of osteogenic genes.