A novel three-dimensional spheroid culture using the ameloblast-lineage cells and the odontoblast-lineage cells for studying maturation in developing tooth germ.

• Project/Area Number: 23592695
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Morphological basic dentistry
• Research Institution: Akita University
• Principal Investigator: KOYOTA Souichi 秋田大学, 学内共同利用施設等, 准教授 (80400480)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: エナメル芽前駆細胞 / スフェロイド培養 / 上皮間葉相互作用 / 歯胚 / 歯胚形成 / 象牙芽前駆細胞 / 細胞分化

Outline of Final Research Achievements

A novel three-dimensional spheroid culture method was developed using the ameloblast-lineage cells (ALC) and the odontoblast-lineage cells (OLC) for studying maturation in developing tooth germ. In the method, the spheroid of ALC and that of OLC fused together so that the tissue of epithelial ALC and that of mesencymal OLC be concentrated and closed expecting to initiate epithelial-mesenchymal interaction. Results from spheroids culture showed that fusions in wide area of the tissue of ALC and that of OLC experienced significant mineralization in whole of construct when compared to their limited small area exhibiting invagination-like processes of epithelial cells. Expression analyses of the genes of differentiation markers and of those in differentiation signal pathways in the spheroid cultures were performed.

Research Products

• [Journal Article] Identification of recurrent SMO and BRAF mutations in ameloblastomas. (2014)
  • Author(s): Sweeney RT, McClary AC, Myers BR, Biscocho J, Neahring L, Kwei KA, Qu K, Gong X, Ng T, Jones CD, Varma S, Odegaard JI, Sugiyama T, Koyota S, Rubin BP, Troxell ML, Pelham RJ, Zehnder JL, Beachy PA, Pollack JR, West RB.
  • Journal Title: Nature Genetics Volume: 46 Issue: 7 Pages: 722-725
  • DOI: 10.1038/ng.2986
  • Peer Reviewed