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Selection of the highly-osteogenic cell population by ghost cytometry for the regeneration of functional bone

Research Project

Project/Area Number 21H03136
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 57060:Surgical dentistry-related
Research InstitutionThe University of Tokyo

Principal Investigator

Hoshi Kazuto  東京大学, 医学部附属病院, 教授 (30344451)

Co-Investigator(Kenkyū-buntansha) 浅輪 幸世  東京大学, 医学部附属病院, 特任講師 (10769912)
疋田 温彦  東京大学, 医学部附属病院, 特任教授 (60443397)
西條 英人  東京大学, 医学部附属病院, 准教授 (80372390)
Project Period (FY) 2021-04-01 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000)
Fiscal Year 2023: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Fiscal Year 2022: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2021: ¥6,630,000 (Direct Cost: ¥5,100,000、Indirect Cost: ¥1,530,000)
Keywords再生医療 / 骨再生 / 細胞分取 / 細胞選択 / 動的ゴーストイメージング / 細胞分離
Outline of Research at the Start

骨形成能の高い骨髄細胞中の細胞集団をsingle cell RNA sequenceあるいは細胞分裂速度の違いを利用して同定する。Ghost Cytometryを用いて、同定した細胞集団を豊富に含む細胞群を分取する方法を確立する。細胞とともに移植する足場として自己組織化ペプチドハイドロゲルを選択し、その濃度や修飾などの仕様を決定する。仕様決定した細胞とペプチドハイドロゲルを組み合わせ、マウス頭蓋骨上皮下に移植し、周囲組織からの圧がかかった状態での骨再生能を評価する。さらにはヒト細胞での細胞分取、さらには大動物での効果検討を行う。

Outline of Final Research Achievements

We tried to establish methods to harvest a highly-osteogenic cell population. When mononuclear cells were harvested from mouse bone marrow cells by the Ficoll gradient centrifugation, the osteogenic ability of the cells were less than those prepared by red blood cell removal. Analysis by flow cytometry indicated that the ratio of CD34-CD73+CD51+cell population was reduced in the cells treated with Ficoll gradient centrifugation, suggesting that the prevention of the loss of this cell population by this method is important. Prior to bone marrow cell experiments, rapid-dividing chondrocytes with high regenerative ability could be concentrated by Ghost cytometry. In addition, by utilizing the dilution of fluorescence dye by cell division, cells were harvested according to the cell division late (high and low), and the osteogenic abilities of both groups were evaluated. Furthermore, several kinds of hydrogels were examined as scaffolds in which cells were to be proliferate.

Academic Significance and Societal Importance of the Research Achievements

骨再生療法の臨床応用が期待されているが、コストの大きさが実用化の障害となっている。本研究では、低コストで優れた骨形成が可能な骨再生法の実現のため、採取した骨髄細胞から骨形成能が高い細胞を選択的に取り出す方法の確立を試みた。また、Ghost cytometryにより再生能の高い軟骨細胞の濃縮が可能であった。この研究の結果は、低コストで質の高い骨再生法の確立に向けて重要な知見を与えるものと考えられる。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Annual Research Report
  • 2021 Annual Research Report
  • Research Products

    (13 results)

All 2024 2023 2022 2021

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 4 results) Presentation (6 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results)

  • [Journal Article] Involvement of Impaired Angiogenesis and Myelosuppression in Antiresorptive-agent Related Osteonecrosis of the Jaw Mouse Model.2023

    • Author(s)
      Igarashi H, Nishizawa S, Miyamoto T, Hikita A, Hoshi K.
    • Journal Title

      Tokai J Exp Clin Med.

      Volume: 48(1) Pages: 22-31

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Subcutaneously Transplanted Fresh Cartilage in Allogeneic and Xenogeneic Immunocompetent Mouse2023

    • Author(s)
      Uto Sakura、Hikita Atsuhiko、Mori Daisuke、Sakamoto Tomoaki、Yano Fumiko、Ohba Shinsuke、Saito Taku、Takato Tsuyoshi、Hoshi Kazuto
    • Journal Title

      Tissue Engineering Part A

      Volume: 29 Issue: 19-20 Pages: 541-556

    • DOI

      10.1089/ten.tea.2023.0052

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Lipoaspirate stored at a constant low temperature by electric control suppresses intracellular metabolism and maintains high cell viability2023

    • Author(s)
      Inaki Ryoko、Sato Yoshihiko、Nakamura Daisuke、Aikawa Yoshiyuki、Takato Tsuyoshi、Hoshi Kazuto、Hikita Atsuhiko
    • Journal Title

      Regenerative Therapy

      Volume: 24 Pages: 662-669

    • DOI

      10.1016/j.reth.2023.11.005

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] miR-92a-3p-inspired shRNA exhibits pro-chondrogenic and chondrocyte protective effects in osteoarthritis treatment through targeting SMAD6/72023

    • Author(s)
      Zheng Chenhuang、Hoshi Kazuto、Hikita Atsuhiko
    • Journal Title

      Journal of Bone and Mineral Metabolism

      Volume: 42 Issue: 1 Pages: 1-16

    • DOI

      10.1007/s00774-023-01474-3

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Involvement of impaired angiogenesis and myelosuppression in antiresorptive-agent related osteonecrosis of the jaw mouse model2023

    • Author(s)
      Hisao Igarashi, Satoru Nishizawa, Takeshi Miyamoto, Atsuhiko Hikita, Kazuto Hoshi
    • Journal Title

      Tokai J Exp Clin Med.

      Volume: 48 Pages: 22-31

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Superior stemness of a rapidly growing subgroup of isolated human auricular chondrocytes and the potential for use in cartilage regenerative therapy2022

    • Author(s)
      Reina Shimizu, Yukiyo Asawa, Makoto Komura, Kazuto Hoshi, Atsuhiko Hikita
    • Journal Title

      Regenerative Therapy

      Volume: 19 Pages: 47-57

    • DOI

      10.1016/j.reth.2021.12.005

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Requirement of direct contact between chondrocytes and macrophages for the maturation of regenerative cartilage.2021

    • Author(s)
      Kengo Kanda, Yukiyo Asawa, Ryoko Inaki, Yuko Fujihara, Kazuto Hoshi, Atsuhiko Hikita
    • Journal Title

      Scientific Reports

      Volume: 11 Issue: 1 Pages: 22476-22476

    • DOI

      10.1038/s41598-021-01437-6

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] ティッシュエンジニアリング技術を利活用した硬組織再生医療2024

    • Author(s)
      星和人
    • Organizer
      第23回日本再生医療学会総会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 口腔科学が牽引する21世紀の医療2023

    • Author(s)
      星和人
    • Organizer
      第77回NPO法人日本口腔科学会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 細胞分裂速度の異なる軟骨細胞の特性評価および純化分離方法の検討2022

    • Author(s)
      清水 玲那, 浅輪 幸世, 能丸 寛子, 古村 眞, 星 和人, 疋田 温彦
    • Organizer
      第76回NPO法人日本口腔科学会学術集会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Label-free machine vision-based cell sorting for tissue engineering2022

    • Author(s)
      H. Nomaru, R. Shimizu, K. Teranishi, Y. Asawa, Y. Kawamura, A. Hikita, K. Hoshi
    • Organizer
      The 28th Annual ISCT Meeting
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Applications of machine vision-based cell sorting for clinical diagnosis and cell therapy2022

    • Author(s)
      Kazuki Teranishi, Hiroko Nomaru, Reina Shimizu, Satoru Akai, Yuri An, Kaoru Komoriya,Yoko Kawamura, Asako Tsubouchi, Yukiyo Asawa, Atsuhiko Hikita, Yusuke Konishi, Kazuto Hoshi, Seiichiro Tabata, Sadao Ota
    • Organizer
      Cyto2022
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 細胞分裂速度の異なる軟骨細胞の特性評価2021

    • Author(s)
      清水 玲那、浅輪 幸世、古村 眞、星 和人、疋田 温彦
    • Organizer
      第21回日本再生医療学会総会
    • Related Report
      2021 Annual Research Report

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Published: 2021-04-28   Modified: 2025-01-30  

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