Abstract: Objective To culture human dental follicle cells in vitro and observe their mineralization characteristics. Methods Human dental follicle tissues were digested with bacterial collagenase and cultured to obtain dental follicle cells. Mineralization characteristics of these cells were identified by immunohistochemistry and reverse transcription (RT)-PCR. Dental follicle cells were incubated with mineralization-inducing agents to observe the formation of mineralized nodules. Results Immunohistochemistry staining revealed expressions of type Ⅰ collagen, type Ⅲ collagen, osteopontin and osteonectin in the cytoplasm of dental follicle cells cultured in vitro. Low levels of bone sialoprotein, osteocalcin and alkaline phosphatase mRNA were also detected by RT-PCR. Mineralized nodules were observed after 20 days of incubation with mineralization- inducing agents with positivity for von-Kossa staining. Conclusion Human dental follicle cells cultured in vitro show some features of osteoblasts or cementoblasts and can be used as the seed cells for periodontal tissue engineering.