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Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability

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Abstract

Objectives

The purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs).

Materials and methods

Human MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1  % fetal bovine serum (low-serum) or α-MEM containing 10  % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation.

Results

Low-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation.

Conclusions

MBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium.

Clinical relevance

These findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.

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Acknowledgements

The authors thank DS Pharma Biomedical for providing the STK2 medium; GC for providing the puncture needle; and the patients in the Department of Regenerative Oral Surgery, Nagasaki University Hospital, for the bone marrow samples. This work was supported by a Grant-in-Aid for Young Researchers (B23792237) and Grants-in-Aid for Scientific Research (C25463007, B22390367, and B26293414) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Oral and Maxillofacial Prosthodontics, Kagoshima University Graduate School of Biomedical Sciences, Kagoshima, Japan

    Fumio Suehiro, Masakazu Ishii & Masahiro Nishimura

  2. Department of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan

    Izumi Asahina

  3. Department of Prosthetic Dentistry, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan

    Hiroshi Murata

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  1. Fumio Suehiro
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  2. Masakazu Ishii
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  3. Izumi Asahina
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  4. Hiroshi Murata
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  5. Masahiro Nishimura
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Correspondence to Masahiro Nishimura.

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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. This article does not contain any studies with animals performed by any of the authors.

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

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Suehiro, F., Ishii, M., Asahina, I. et al. Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability. Clin Oral Invest 21, 2709–2719 (2017). https://doi.org/10.1007/s00784-017-2073-7

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  • DOI: https://doi.org/10.1007/s00784-017-2073-7

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