Comparison of Primary Ameloblast and Ameloblast-like Cell Line LS8 for Enamel-specific Gene Expression With High-fluoride

Objectives: In this study we investigated if ameloblast-like cell line (LS8), a substitution of ameloblast in vitro experiment, has the same reaction with primary ameloblast when exposing to the high-fluoride.
Methods: （1）The tooth germs were separated from 4 days old SD rats and cleaned by PBS with. The tissues were added in collagenase for 2 hours. Then the tissues were incubated repeatedly with trypsin. the cells were suspended in DMEM with 15% heat inactivated fetal bovine serum (FBS) and were cultured . After 48 hours，fibroblast-like cells were removed by trypsin. The ameloblast-like cell line LS8 cells were cultured in DMEM with 10% FBS. (2) The primary ameloblast and LS8 cells were processed for determination of cell viability and the mRNA expression of FoxO1，KLK4 and Amtn after fluoride exposure using MTT and real-time RT-PCR.
Results: （1）MTT revealed: compared with the control group (0 mmol/L NaF group), cell viability of both primary ameloblasts and LS8 cells decreased gradually in the two experiment groups in a dependent manner with the concentration of NaF (p < 0.05）. (2) Real-time RT-PCR showed that Amtn,KLK4 and FoxO1 mRNA expression decreased in both primary ameloblast and LS8 cells after fluoride expose.
Conclusions: LS8 cells showed the same reaction with primary ameloblasts in cell viability and the mRNA expression of Amtn,KLK4 and FoxO1 after exposing to fluoride.