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Isolation and characterization of dental follicle–derived Hertwig’s epithelial root sheath cells

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Abstract

Objectives

The aim of this study was the isolation and characterization of dental follicle–derived Hertwig’s epithelial root sheath cells (DF-HERSCs).

Materials and methods

DF-HERSCs were isolated from dental follicle (DF)–derived single-cell suspensions. Their epithelial phenotypes were analyzed by Western blotting, polymerase chain reaction (PCR), and quantitative polymerase chain reaction (qPCR). Epithelial-mesenchymal transition (EMT) was induced in DF-HERSCs by treatment with transforming growth factor-β (TGF-β) or fetal bovine serum (FBS)–added medium. Characteristics of DF-HERSCs were compared with normal human oral keratinocytes (NHOKs) and normal human epidermal keratinocytes (NHEKs). Osteogenic differentiation and mineralization of DF-HERSCs were analyzed by alkaline phosphatase (ALP) and Alizarin red staining. All experiments were conducted in triplicate.

Results

Primary DF-HERSCs were isolated from DF. Epithelial phenotypes of DF-HERSCs were confirmed by morphological and Western blot analysis. PCR results demonstrated that the origin of DF-HERSCs was neither endothelial nor hematopoietic. Enamel matrix derivative (EMD)–associated genes were not expressed in DF-HERSCs. Treatment with TGF-β and FBS-added medium triggered the progression of EMT in DF-HERSCs. The acquired potency of differentiation and mineralization was shown in EMT-progressed DF-HERSCs.

Conclusions

DF contains putative populations of HERSC, named DF-HERSC. DF-HERSCs shared common characteristics with NHOKs and NHEKs.

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Acknowledgments

Professor Dong-Mok Ryu kindly provided tissues for primary culture.

Funding

This study was supported by the grants from the National Research Foundation of Korea (NRF) funded by the Korean government (NRF-2017R1C1B2008406) and Kyung Hee University in 2017(KHU-20170853).

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Authors and Affiliations

  1. Anesthesia and Pain Research Institute, School of Medicine, Yonsei University, Seoul, South Korea

    Ju Eun Oh

  2. Department of Conservative Dentistry, School of Dentistry, Kyung Hee University, Seoul, South Korea

    Jin-Kyu Yi

  3. Department of Conservative Dentistry, Kyung Hee University Dental Hospital at Gangdong, 892, Dongnam-Ro, Gangdong-Gu, Seoul, 05278, South Korea

    Jin-Kyu Yi

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  1. Ju Eun Oh
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Correspondence to Jin-Kyu Yi.

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The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Clinical relevance

Scientific evidence of the roles of DF-HERSCs, especially during cementogenesis, might be utilized for the development of strategies aimed the endodontic regeneration and periodontal re-attachment. Procuring human DF-HERSCs can facilitate HERS-associated studies.

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Oh, J.E., Yi, JK. Isolation and characterization of dental follicle–derived Hertwig’s epithelial root sheath cells. Clin Oral Invest 25, 1787–1796 (2021). https://doi.org/10.1007/s00784-020-03481-4

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  • DOI: https://doi.org/10.1007/s00784-020-03481-4

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