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Establishment of clinical culture method for human dental pulp stem cells under the xeno-free culture condition.

Research Project

Project/Area Number 19K21403
Project/Area Number (Other) 18H06317 (2018)
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeMulti-year Fund (2019)
Single-year Grants (2018)
Review Section 0907:Oral science and related fields
Research InstitutionThe Nippon Dental University

Principal Investigator

Mochizuki Mai  日本歯科大学, 生命歯学部, 助教 (90821934)

Project Period (FY) 2018-08-24 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords無血清培養 / 歯髄幹細胞 / 間葉系幹細胞 / 細胞培養 / 再生医療
Outline of Research at the Start

これまで我々は、無血清培養下において歯髄組織からの幹細胞の分離と大量培養に成功し、同時に無血清培養における歯髄幹細胞が、過度な培養によって多層化して細胞死が生じることを突き止めた。
本研究は、無血清培養による臨床的培養法の確立をゴールとし、過度な培養による多層化と細胞死の原因を明らかにするため、歯髄幹細胞が呈する特異的な接着形態に着目し、その細胞接着機構の解明を目的とする。すなわち、無血清培養液による細胞周囲の環境によって生じた細胞形態に基づき、その細胞形態によって生じる細胞内シグナル経路を突き止め、過度な培養で細胞死をまねく歯髄幹細胞の多層化と細胞増殖能が破綻するメカニズムの解明を試みる。

Outline of Final Research Achievements

We previously reported that a xenogeneic serum-free culture medium (XFM) is preferable over fetal bovine serum-containing culture medium for ex vivo expansion of human dental pulp-derived stem cells (hDPSCs). However, we observed that cells grown in XFM frequently undergo apoptosis when they form multilayered cell structures upon reaching over-confluence. Therefore, we focused on optimizing the XFM culture system to avoid the undesirable death of hDPSCs.
This study demonstrated that type 1 collagen coating of the cultureware promoted hDPSC adhesion and proliferation. Furthermore, this coating prevented cell death due to multilayered cell growth in XFM conditions. The results provide further insights into the safe and reliable handling of somatic stem cell cultures.

Academic Significance and Societal Importance of the Research Achievements

本研究は、がん化リスクのないヒト歯髄幹細胞の再生医療応用に向け、細胞治療に有用な培養法の確立を目指し、無血清培養における効果的な歯髄幹細胞培養に貢献する知見を提供した。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • Research Products

    (10 results)

All 2020 2019 2018

All Presentation (10 results) (of which Int'l Joint Research: 2 results,  Invited: 1 results)

  • [Presentation] Type 1 collagenを用いたヒト歯髄幹細胞の臨床的培養法の確立2020

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      第19回日本再生医療学会総会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 再生医療に向けたヒト歯髄幹細胞における臨床的培養法の確立2019

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      第18回日本再生医療学会総会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 再生医療のための無血清培養およびフィブロネクチンコーティングによるヒト歯髄幹細胞の分離と同定2019

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      令和元年度日本歯科大学歯学会大会・総会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 異種血清非存在下におけるヒト歯髄幹細胞の分離・同定とDMSOフリー凍結保存の検証~再生医療に向けた臨床的培養法の確立~2019

    • Author(s)
      望月真衣
    • Organizer
      第19回日本抗加齢医学会総会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Effects of type I collagen on human dental pulp stem cells grown in xenogeneic serum-free medium for establishment of a practical culture method2019

    • Author(s)
      Mochizuki M, Nakahara T
    • Organizer
      17th Annual Meeting International Society for Stem Cell Research
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 無血清培養によるヒト歯髄幹細胞の大量培養に向けたI型コラーゲンの可能性~安全を約束する臨床的培養法の確立~2019

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      日本組織培養学会 第92回大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 臨床的歯根再生に向けた歯根膜幹細胞の特性評価2019

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      第37回日本ヒト細胞学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ゼノフリー無血清培養下におけるⅠ型コラーゲンの影響~ヒト歯髄幹細胞の臨床的培養法の確立~;Effect of type 1 collagen under xeno-free culture condition for the establishment of the clinical culture method using human dental pulp stem cells2019

    • Author(s)
      望月真衣, 中原 貴
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Effects of type i collagen on the growth of human dental pulp stem cells in a xenogeneic serum-free medium for the establishment of a practical culture method2019

    • Author(s)
      Mai Mochizuki, Taka Nakahara
    • Organizer
      17th Annual Meeting International Society for Stem Cell Research
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] ゼノフリー無血清培養におけるヒト歯髄幹細胞の細胞死の究明~臨床的培養法の確立に向けて~2018

    • Author(s)
      望月真衣、中原 貴
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report

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Published: 2018-08-27   Modified: 2024-03-26  

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