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Development of Optimal Cell Culture Method with Morphological Quality Assessment by DNA Damage

Research Project

Project/Area Number 26293422
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypePartial Multi-year Fund
Section一般
Research Field Dental engineering/Regenerative dentistry
Research InstitutionNational Center for Geriatrics and Gerontology

Principal Investigator

NAKASHIMA Misako  国立研究開発法人国立長寿医療研究センター, 幹細胞再生医療研究部, 部長 (20207773)

Co-Investigator(Kenkyū-buntansha) 山本 徳則  名古屋大学, 医学(系)研究科(研究院), 准教授 (20182636)
村上 真史  国立研究開発法人国立長寿医療研究センター, その他部局等, その他 (30614531)
栗田 賢一  愛知学院大学, 歯学部, 教授 (40133483)
兼子 智  東京歯科大学, 歯学部, 講師 (40214457)
エンドラワン プラタマ  国立研究開発法人国立長寿医療研究センター, 幹細胞再生医療研究部, 流動研究員 (10783491)
Research Collaborator IOHARA Koichiro  
AHMED Nermeen  
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000)
Fiscal Year 2016: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2015: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2014: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000)
Keywords歯髄幹細胞 / マルチガスインキュベータ / 微小重力培養 / 品質検査 / DNA損傷 / DNA障害 / 品質評価 / DNA損傷 / マルチインキュベーター / マルチガスインキュベータ / DNA傷害 / DNA傷害
Outline of Final Research Achievements

The present study aimed at development of the optimal culture method and quality control and assurance method for dental pulp stem cells (DPSCs). At first, the morphological quality assessment of genomic DNA damage was performed by Single-Cell Pulsed-Field Gel Electrophoresis (SCPFGE) as a quality control and assurance method. The enzymatic or physical detachment to get a single cell suspension, however, caused a nonspecific DNA damage. Next, the optimal hypoxic condition for DPSCs was investigated in a tri-gas incubator for stable culture. The proliferation rate, stem cell properties and trophic effect were the highest in 5% O2 culture. Furthermore, three-dimensional microgravity devise were used. There was, however, little difference in the proliferation rate, cell stability and quality in microgravity compared with normal gravity in only one passage of culture. Further three passages of culture under microgravity resulted in little difference in these from normal gravity.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Annual Research Report
  • 2014 Annual Research Report
  • Research Products

    (17 results)

All 2016 2015 2014

All Journal Article (5 results) (of which Peer Reviewed: 5 results,  Open Access: 3 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 8 results)

  • [Journal Article] Effects of extracellular pH on dental pulp cells in vitro2016

    • Author(s)
      Hirose Y, Yamaguchi M, Kawabata S, Murakami M, Nakashima M, Gotoh M, Yamamoto T.
    • Journal Title

      J Endod.

      Volume: 42(5) Pages: 735-741

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The effects of hypoxia on the stemness properties of human dental pulp stem cells (DPSCs)2016

    • Author(s)
      Ahmed NE, Murakami M, Kaneko S, Nakashima M.
    • Journal Title

      Sci Rep.

      Volume: 6 Issue: 1 Pages: 35476-35476

    • DOI

      10.1038/srep35476

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Effects of extracellular pH on Dental Pulp Cells in Vitro2016

    • Author(s)
      Hirose, Y., Yamaguchi, M., Kawabata, S., Murakami, M., Nakashima, M., Gotoh, M., Yamamoto, T.
    • Journal Title

      Journal of endodontics

      Volume: 印刷中 Issue: 5 Pages: 735-741

    • DOI

      10.1016/j.joen.2016.01.019

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Mechanical forces induce odontoblastic differentiation of mesenchymal stem cells on three-dimensional biomimetic scaffolds.2014

    • Author(s)
      Miyashita S, Ahmed N E, Murakami M, Iohara K, Yamamoto T, Horibe H, Kurita K, Takano-Yamamoto T, Nakashima M
    • Journal Title

      J Tissue Eng Regen Med

      Volume: 11 Issue: 2 Pages: 434-446

    • DOI

      10.1002/term.1928

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Tissue engineering-based strategies for regenerative endodontics.2014

    • Author(s)
      Albuquerque M T P, Valera M C, Nakashima M, Nor J.E, Bottino M C
    • Journal Title

      J Dent Res

      Volume: 93 Pages: 1222-1231

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 歯髄・象牙質再生治療の現状と未来2016

    • Author(s)
      中島美砂子
    • Organizer
      第23回日本歯科医学会総会
    • Place of Presentation
      福岡
    • Year and Date
      2016-10-22
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Optimizing oxygen conditions for expansion of human dental pulp stem cells (DPSCs) under controlled oxygen concentration2016

    • Author(s)
      Ahmed NE, Murakami M, Kaneko S, Nakashima M
    • Organizer
      International Association for Dental Research (IADR) Pulp Biology and Regeneration Group (PBRG) Symposium 2016
    • Place of Presentation
      名古屋
    • Year and Date
      2016-06-27
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 歯の延命化による健康長寿を目指した歯髄・象牙質再生治療法の現状と展望2015

    • Author(s)
      中島美砂子
    • Organizer
      シンポジウム「健康長寿と再生医療」日本学術会議歯学委員会主催、日本歯科医学会・日本歯学系学会協議会共催
    • Place of Presentation
      東京
    • Year and Date
      2015-12-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] Dental pulp stem cells mobilized with granulocyte colony-stimulating factor for total pulp regeneration: Phase 1 clinical trial2015

    • Author(s)
      Nakashima M
    • Organizer
      Proceedings of International Conference on Clinical Trials
    • Place of Presentation
      オーランド
    • Year and Date
      2015-07-27
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 歯の延命化を目指した歯髄幹細胞自家移植による歯髄再生治療法の臨床研究2015

    • Author(s)
      中島美砂子
    • Organizer
      第33回日本骨代謝学会学術集会
    • Place of Presentation
      東京
    • Year and Date
      2015-07-24
    • Related Report
      2015 Annual Research Report
  • [Presentation] Dental Pulp Stem Cells Mobilized with Granulocyte Colony-Stimulating Factor for Total Pulp Regeneration Clinical Trial2015

    • Author(s)
      Nakashima M
    • Organizer
      5th Research Week International Symposium of Oral Medicine College of Oral Medicine
    • Place of Presentation
      台北
    • Year and Date
      2015-05-16
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 自己歯髄組織由来幹細胞を用いた抜髄後歯髄再生療法の臨床研究2015

    • Author(s)
      中島美砂子
    • Organizer
      第14回日本再生医療学会総会 シンポジウム
    • Place of Presentation
      横浜
    • Year and Date
      2015-03-21
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] シリコーン膜を用いた加圧刺激によるヒトiPS細胞の象牙芽細胞への分化誘導2015

    • Author(s)
      村上真史、宮下俊郎、Nermeen Kandel、庵原耕一郎、中島美砂子
    • Organizer
      第14回日本再生医療学会総会
    • Place of Presentation
      横浜
    • Year and Date
      2015-03-19
    • Related Report
      2014 Annual Research Report
  • [Presentation] 自己歯髄幹細胞を用いた歯髄再生治療法の臨床研究2014

    • Author(s)
      中島美砂子
    • Organizer
      第141回日本歯科保存学会秋季学術大会シンポジウム
    • Place of Presentation
      山形
    • Year and Date
      2014-10-31
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 歯髄再生医療と医工学2014

    • Author(s)
      庵原耕一郎、宮下俊郎、中島美砂子
    • Organizer
      応用物理学会東海支部・基礎セミナー バイオメディカルエンジニアリング
    • Place of Presentation
      名古屋
    • Year and Date
      2014-09-05
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] Clinical evidences of stem cell-based therapies in regenerative endodontics. Tissue engineering strategies for regenerative endodontics2014

    • Author(s)
      Nakashima M
    • Organizer
      The International Association for Dental Research (IADR)
    • Place of Presentation
      南アフリカ Cape Town
    • Year and Date
      2014-06-26
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] Dental pulp stem cell biology and regeneration for clinical trial2014

    • Author(s)
      Nakashima M
    • Organizer
      The Second International Conference on Dental and Craniofacial Stem Cells (ICDCSC)
    • Place of Presentation
      中国 成都
    • Year and Date
      2014-04-12
    • Related Report
      2014 Annual Research Report
    • Invited

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Published: 2014-04-04   Modified: 2019-12-27  

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