Objective To establish a method for culturing dental follicle cells of rat and observe the growth characteristics of the cultured cellsin vitro.Methods Dental follicle and associated enamel organs were dissected from the first and second man- dibular molars of 6-7-day-old rats, and then culturedin vitro. Purified dental follicle cells derived fromthe third orthe fourth pas- sage cells were utilized in the following experiments. The shape and ultrastructure of dental follicle cellswere observed by light-mi- croscopy and transmission electron microscopy. Immunocytochemistrywas used to detect the expression of vimentin, typeⅠcolla- gen and fibronectin.Results The cultured cells were fibroblastic in shape, elongate with processes, and transmission electron microscopy revealed that they contained electron-dense granules, an abundant rough endoplasmic reticulum, but did not form desmosomes. Vimentin, typeⅠcollagen and fibronectinwere present in dental follicle cell.Conclusion The dental follicle cells of rat could be successfully culturedin vitroand the cultured cells had the same characteristics of dental follicle cells of normal rat.