Abstract
The biochemical study on the dentinogenesis of unerupted tooth germ including development, eruption and mastication is of great value to all areas of dentistry, especially to the pedodontic practice. In this study, we contrived a simple procedure for cell culture from a bovine dental sac as the first step of cell characterization.
A fresh tooth germ was dissected from a 2-year-old bovine jaw. Simple pieces of the tooth germ were placed between the bottom of a 2 cm2 well and a 15 mm thermanox coverslip in D-MEM, containing 2 mg FCSP/ml,50μg ascorbic acid/ml and penicillin/streptomycin, using the Kawase and Saito method modified from Brunette. The dental sac cells of each stages were grown out of an explant and then subcultured by trypsinization. The cultures were maintained at 37°C and treated with or without 5×10-9 M 1,25(OH)2D3 and 5×10-8M 24,25 (OH)2D3.
Durin g the explantation, the cells of stage I (inner) and stage III (outer)migrated out actively. Morphologically, the migrated cell population was extremely heterogeneous. Nevertheless, the shape of the fibroblast-like cells, which were mainly grown with a day of primary culture, did not change for at least several passages. The growth of subcultured cells of each stages was inhibited by 1,25(OH)2D3, but was not inhibited by 24,25(OH)2D3 at any stages.
Therefore, cultures of dental sacs were able to be demonstrated by means of the outgrowth migration from explants and characterized with respect to the effect of vitamin D3.
