| Co-Investigator(Kenkyū-buntansha) |
TSUNODA Akira Kanagawa Dental College, Department of Operative Dentistry and Endodontics, Lecturer, 歯学部, 講師 (70236933)
YAMAUCHI Masato Kanagawa Dental College, Department of Orthodontics, Assistant Professor, 歯学部, 助手 (30230311)
KIYONO Thoru National Cancer Center Research Institute, Virology Division, Chief, ウイルス部, 部長 (10186356)
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| Research Abstract |
Dental follicle is the fibrous tissue that surrounds the developing tooth germ, and it is believed to contain progenitors for cementoblasts, periodontal ligament cells and osteoblasts. To investigate mechanisms of cementogenesis, bovine dental follicle cells (BDFC) were obtained from bovine tooth germs In culture, BDFC exhibited low levels of alkaline phosphatase activity and expressed mRNA for osteopontin (OP) and type I collagen (COLI), as well as low levels of osteocalcin (OC) mRNA. To elucidate the differentiation capacity of BDFC in vivo, cells were transplated into severe combined immunodeficiency (SCID) mice and analyzed after 4 weeks.
Transplanted BDFC formed fibrous tissue and cementum-like matrix, which stained positive for anti-cementum attachment protein (CAP) monoclonal antibody (3G9), and expressed mRNA for OC, OP, COLI and bone sialoprotein (BSP). These results indicate that cementoblast progenitors are present in BDFC. To further analysis of cementoblast progenitor, attempt was made to immortalize BDFC. BDFC was immortalized by combination of human papilloma virus type 16 E6E7 and telomerase reverse transcriptase subunit (hTERT) with retrovirus transfer. After gene transfer, BDFC was successfully immortalized, they retained same morphology and cell proliferating activity even when the cells prolonged culture up to PD100. From these findings, cementoblast progenitor cell line seems to isolate from immortalize BDFC. To achieve this goal, we will plan to establish cementoblast progenitor cell line using single cell based analysis.
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