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Isolation and characterization of SSEA-4-positive subpopulation of human deciduous dental pulp cells

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Abstract

Objectives

It is well accepted that stage-specific embryonic antigen (SSEA)-4 is an antigen that is useful to isolate adult stem cells analogous to embryonic stem cells. Therefore, in the present study, we investigated whether SSEA-4 can also be used as a marker to identify human deciduous dental pulp (D-DP) stem cells.

Materials and methods

Intact deciduous teeth were collected from healthy patients who were undergoing orthodontic treatment at Okayama University Hospital. Immunofluorescence analysis, flow cytometric analysis, and multilineage differentiation assay were performed to characterize SSEA-4+ D-DP cells.

Results

The D-DP cells had the characteristics of mesenchymal stem cells (MSCs), namely plastic adherence, specific surface antigen expression, and multipotent differentiation potential. SSEA-4 expression was detected in D-DP cells in vitro and ex vivo samples. A flow cytometric analysis demonstrated that 21.2 % of the D-DP cells were positive for SSEA-4. The SSEA-4+ clonal D-DP cells showed multilineage differentiation potential toward adipocytes, osteoblasts, and chondrocytes in vitro. In fact, 26.1 % (6/23) of the SSEA-4+ clonal D-DP cells showed adipogenic potential, 91.3 % (21/23) showed osteogenic potential, 91.3 % (21/23) showed chondrogenic potential, and 87.0 % (20/23) showed both osteogenic and chondrogenic potential.

Conclusions

Thus, the majority of SSEA-4+ D-DP cells had the potential for multilineage differentiation. Hence, SSEA-4 appears to be a specific marker that can be used to identify D-DP stem cells.

Clinical relevance

SSEA-4+ D-DP cells appear to be a promising source of stem cells for regenerative therapy.

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Acknowledgments

The present study was supported by Grants-in-Aid for Scientific Research (to N. Kawanabe [24593091] and T. Yamashiro [24249093]) from the Japan Society for the Promotion of Science, Japan.

Author contribution

N. Kawanabe contributed to the conception and design of the study, obtaining financial support for the study, the collection and/or assembly of the data, the data analysis and interpretation, the writing of the manuscript, and the final approval of the manuscript. H. Fukushima contributed to the conception and design of the study, the collection and/or assembly of the data, the data analysis and interpretation, and the writing of the manuscript. Y. Ishihara performed the confocal laser microscopic analysis. T. Yamashiro contributed to the conception and design of the study, obtaining financial support, the data analysis and interpretation, and the writing of the manuscript.

Conflict of interest

The authors declare no potential conflicts of interest with respect to the authorship and/or publication of this article.

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Authors and Affiliations

  1. Department of Orthodontics, Okayama University Hospital, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan

    Noriaki Kawanabe & Takeshi Yanagita

  2. Department of Orthodontics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

    Hiroaki Fukushima, Yoshihito Ishihara & Hiroshi Kurosaka

  3. Department of Orthodontics and Dentofacial Orthopedics, Graduate School of Dentistry, Osaka University, Osaka, Japan

    Takashi Yamashiro

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  1. Noriaki Kawanabe
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  2. Hiroaki Fukushima
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  3. Yoshihito Ishihara
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  4. Takeshi Yanagita
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  5. Hiroshi Kurosaka
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  6. Takashi Yamashiro
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Correspondence to Noriaki Kawanabe.

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Kawanabe, N., Fukushima, H., Ishihara, Y. et al. Isolation and characterization of SSEA-4-positive subpopulation of human deciduous dental pulp cells. Clin Oral Invest 19, 363–371 (2015). https://doi.org/10.1007/s00784-014-1260-z

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  • DOI: https://doi.org/10.1007/s00784-014-1260-z

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