title: Establishment and characterization of a spontaneously immortalized mouse ameloblast-lineage cell line.
abstract: Tooth development was cooperatively regulated by the epithelial ameloblasts and mesenchymal odontoblasts. Ameloblasts secrete enamel matrix, critical for enamel formation. While there are several reports about establishment of immortalized ameloblast-like cells by introducing viral oncogene, we tried to establish a spontaneously immortalized ameloblast-lineage cell line, maintaining the cell type specific character, including the ability to induce in vitro bio-mineralization. The established cell line (ameloblast-lineage cell; ALC) maintained the expression of several ameloblast specific genes (Amelogenin, Tuftelin, and Enamelin) in long-term culture. They formed calcified nodules after the induction by medium switching from SMEM to DMEM, having high-level alkaline-phosphatase activity. The size and number of calcified nodule formation were enhanced by TGF-beta treatment. Six weeks after sub-cutaneous implantation of ALC to athymic nude mice, we ectopically observed enamel epithelium like structure formation, chondrogenesis, and calcification. These data indicate that ALC is a useful experimental tool to analyze ameloblast character.
authors.affiliation: Department of Biochemistry, Akita University School of Medicine, Hondo 1-1-1, Akita 010-8543, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Nakata
authors.forename: Takashi
authors.initials: T
authors.lastname: Kameda
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Nagai
authors.forename: Kenji
authors.initials: K
authors.lastname: Ikegami
authors.forename: Yuqing
authors.initials: Y
authors.lastname: Duan
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Terada
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Sugiyama
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000565:Ameloblasts / Q000166:cytology*; D053523:Amelogenin; D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D019070:Cell Lineage; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D003746:Dental Enamel Proteins / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014070:Tooth / Q000166:cytology*; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D053523:Amelogenin; C507200:Amelx protein, mouse; D018076:DNA, Complementary; D003746:Dental Enamel Proteins; D016212:Transforming Growth Factor beta; C038456:tuftelin; D000469:Alkaline Phosphatase; D002118:Calcium