Establishment of Immortal Dog Dental Pulp Cell Line and Analysis of Its Characteristics

DOI 20 References
  • HANDA Keisuke
    Division of Clinical Cariology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido
  • KOIKE Toshiyuki
    Division of Clinical Cariology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido
  • KIYONO Tohru
    Virology Division, National Cancer Center Research Institute
  • SAITO Takashi
    Division of Clinical Cariology and Endodontology, Department of Oral Rehabilitation, School of Dentistry, Health Sciences University of Hokkaido

Bibliographic Information

Other Title
  • 不死化イヌ歯髄細胞株の樹立とその特性の解析

Search this article

Description

Since normal cells in a primary cell culture possess a Hyflick limit, which is the number of times cells divide before they stop due to the telomere reaching a critical length, it is difficult to analyze odontoblasts in vitro sufficiently. As a result, little is known about the mechanism of odontoblasts differentiation; extending the life time of odontoblasts may enable a detailed analysis of them in vitro. The purposes of this study were to establish an immortal dog dental pulp cell line by the gene transfection of human telomere reverse transcriptase (hTERT), human papilloma virus (HPV) E6 and E7 into dog dental pulp cells (DDP) in vitro and to analyze its characteristics. First, we obtained an immortal dog dental pulp cell line, DDP (E6E7/TERT) by infection of retrovirus recombined with hTERT, HPV E6 and E7 to DDP derived from dog anterior teeth. Next, ALPase staining and Alizarin Red staining were carried out to monitor differentiation and mineralization in DDP (E6E7/TERT) in vitro. In both stainings, positive staining was defined in DDP (E6E7/TERT) on day 6. Moreover, the expressions of hard tissue related genes (osteopontin, type I collagen, Runx2) were observed by the RT-PCR method. These results indicated that DDP (E6E7/TERT) was immortalized while maintaining the same characteristics as normal cells, suggesting the immortalization system used in this study is effective for the immortalization of dog dental pulp cells. Also, it was suggested that DDP (E6E7/TERT) might be a useful tool for clarifying the mechanism of odontoblasts differentiation and developing a novel pulp capping agent.

Journal

References(20)*help

See more

Related Projects

See more

Keywords

Details 詳細情報について

Report a problem

Back to top