OBJECTIVETo establish a method for isolating and culturing mouse dental follicle cells and to study the phenotype characteristics of dental follicle cells.

METHODSMandibular first molars from 9 day old Balb/c mice were digested with 1% trypsin, subsequently, and the dental follicle was enucleated from the tooth germ and cultured. The shape and ultrastructural appearance of dental follicle cells were observed under phase-contrast microscope and scanning electron microscope (SEM). Immunocytochemistry was used to detect the expression of alkaline phosphatase (ALP), bone sialoprotein (BSP) and osteopontin (OPN).

RESULTSThree types of cells were isolated: some were cuboidal/polygonal; some were elongated, spindle-shaped, fibroblast-like cells; and a minor, third cell type was very thin and elongated. The cytoplasm of the first two cell types was filled with abundant granules. The cells were pleomorphism under SEM and had many filipodia and microvilli. According to whether there were filaments overlying the surface, the cells could be divided into two subtypes. Some but not all follicle cells expressed ALP, BSP and OPN.

CONCLUSIONThe cultured dental follicle cells consisted of several cell phenotypes and had the potential of differentiating into cementoblasts, periodontal ligament fibroblasts and osteoblasts.