[P3-3-17] Isolation of odontoblast and ameloblast progenitors using Dspp-GFP or Amelx-tdTomato mice that can label odontoblasts and ameloblasts fluorescently
Keywords:発生・分化、象牙芽細胞、エナメル芽細胞
We have established Dspp-GFP and Amelx-tdTomato mice by inserting green or red fluorescent protein genes into the Dspp and Amelx loci, respectively, to identify odontoblasts and ameloblasts using fluorescence as an indicator. We have previously reported that co-culture of embryonic dental mesenchyme cells from normal mice and Dspp-GFP mice with embryonic dental epithelium from control mice revealed the presence of GFP-negative odontoblast precursors that became GFP-positive after tooth organ culture. Similarly, we reported that co-culture of embryonic dental epithelial cells from normal mice and Amelx-tdTomato mice with embryonic dental mesenchymal cells from normal mice resulted in the presence of tdTomato-negative ameloblast precursors that became tdTomato-positive after tooth organ culture. In the present study, to isolate odontoblast progenitor cells, we used tooth germs from 13.5- or 14.5-day-old embryos and confirmed the expression of cell surface molecules mainly by immunohistochemistry staining or flow cytometry, and isolated a population of odontoblast progenitor cells in CD90-positive or CD90-negative fractions that were GFP positive from GFP negative after tooth organ culture. Similarly, to isolate ameloblast progenitor cells, we used tooth germs from 13.5- or 14.5-day-old embryos to confirm the expression of cell surface molecules mainly by immunohistochemistry staining or flow cytometry. We report to attempt for the isolation of a cell population of ameloblast progenitor cells that were tdTomato positive from tdTomato negative after tooth organ culture.