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Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo

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Abstract

The dental follicle is a mesenchymal tissue that surrounds the developing tooth germ. During tooth root formation, periodontal components, viz., cementum, periodontal ligament (PDL), and alveolar bone, are created by dental follicle progenitors. Here, we report the presence of PDL progenitors in mouse dental follicle (MDF) cells. MDF cells were obtained from mouse incisor tooth germs and immortalized by the expression of a mutant human papilloma virus type 16 E6 gene lacking the PDZ-domain-binding motif. MDF cells expressing the mutant E6 gene (MDFE6-EGFP cells) had an extended life span, beyond 150 population doublings (PD). In contrast, normal MDF cells failed to proliferate beyond 10 PD. MDFE6-EGFP cells expressed tendon/ligament phenotype-related genes such as Scleraxis (Scx), growth and differentiation factor-5, EphA4, Six-1, and type I collagen. In addition, the expression of periostin was observed. To elucidate the differentiation capacity of MDFE6-EGFP cells in vivo, the cells were transplanted into severe combined immunodeficiency mice. At 4 weeks, MDFE6-EGFP cell transplants had the capacity to generate a PDL-like tissue that expressed periostin, Scx, and type XII collagen and the fibrillar assembly of type I collagen. Our findings suggest that MDFE6-EGFP cells can act as PDL progenitors, and that these cells may be a useful research tool for studying PDL formation and for developing regeneration therapies.

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Acknowledgement

We are grateful to Dr. Masato Yamauchi for his advice and discussions during the course of this work.

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Authors and Affiliations

  1. Department of Medicine, Division of Operative Dentistry and Endodontics, Kanagawa Dental College, 82 Inaoka-cho, Yokosuka, Kanagawa, 238-8580, Japan

    T. Yokoi, M. Saito, K. Kosaka, E. Nishida, T. Tsubakimoto & T. Teranaka

  2. Department of Periodontology, School of Dentistry, Aichi-gakuin University, Nagoya, Aichi, Japan

    T. Yokoi, E. Nishida & T. Noguchi

  3. Oral Health Science Research Center, Kanagawa Dental College, Yokosuka, Kanagawa, Japan

    M. Saito

  4. Virology Division, National Cancer Research Institute, Tokyo, Japan

    T. Kiyono

  5. Department of Molecular Craniofacial Embryology,Graduate School, Tokyo Medical and Dental University, Tokyo, Japan

    S. Iseki & K. Eto

  6. Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, Suita, Osaka, Japan

    H. Harada

Authors
  1. T. Yokoi
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  2. M. Saito
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  3. T. Kiyono
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  4. S. Iseki
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  5. K. Kosaka
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  6. E. Nishida
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  7. T. Tsubakimoto
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  8. H. Harada
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  9. K. Eto
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  10. T. Noguchi
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  11. T. Teranaka
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Corresponding author

Correspondence to M. Saito.

Additional information

This work was supported by a Grant-in Aid for the High-Tech Research Center Project from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the AGU High-Tech Research Center Project, the 2003-Multidisciplinary Research Project from MEXT, and grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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Yokoi, T., Saito, M., Kiyono, T. et al. Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo. Cell Tissue Res 327, 301–311 (2007). https://doi.org/10.1007/s00441-006-0257-6

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  • DOI: https://doi.org/10.1007/s00441-006-0257-6

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