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Genetically-encoded calcium indicators such as G-GECO are useful for studying Ca 2+ responses during long-term processes. In this study, we employed a lentiviral vector and established a rat dental epithelial cell line that stably expressed G-GECO (SF2-G-GECO). Ca 2+ imaging analysis under cell culture conditions revealed that SF2-G-GECO cells exhibited spontaneous Ca 2+ responses, which could be classified into the following three major patterns depending on the cell density: localized Ca 2+ responses at cell protrusions at a low density, a cell-wide spread of Ca 2+ responses at a medium density, and Ca 2+ responses in clusters of 3-20 cells at a high density. The P2Y receptor inhibitor suramin (10 μM), the ATP-degrading enzyme apyrase (5 units/mL), and the fibroblast growth factor (FGF) receptor inhibitor FIIN-2 (1 μM) decreased the frequency of spontaneous Ca 2+ responses. These results indicate that ATP and FGF are involved in the spontaneous Ca 2+ responses. SF2 cells differentiate into ameloblasts via interactions with mesenchymal cells. Therefore, SF2-G-GECO cells are expected to be a useful tool for studying the functions of Ca 2+ responses in regulating gene expression during tooth development.

Spontaneous calcium responses of SF2 rat dental epithelial cells stably expressing the calcium sensor G-GECO