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Studies on cryopreservation mechanism of intracellular trehalose introduced through trehalose transporter

Research Project

Project/Area Number 17K18834
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Research Field Fluid engineering, Thermal engineering, and related fields
Research InstitutionHokkaido University

Principal Investigator

UCHIDA TSUTOMU  北海道大学, 工学研究院, 准教授 (70356575)

Research Collaborator Kikawada Takahiro  農業・食品産業技術総合研究機構, 主任研究員
Takeya Satoshi  産業技術総合研究所, 主任研究員
Takeya Kei  名古屋大学, 工学研究科, 講師
Project Period (FY) 2017-06-30 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2017: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Keywordsトレハロース / 凍結保存 / トレハロース輸送体 / 細胞内水 / ガラス化 / 氷結晶 / CHO-K1細胞 / TRET1 / 緩慢凍結 / 熱工学 / 流体工学 / 細胞・組織
Outline of Final Research Achievements

In this study, we aimed to clarify the mechanism of the high cryoprotective effect of intracellular trehalose experimentally. Since trehalose is not transported into the cell spontaneously, we used two CHO-K1 cells as the model cell, one of which is expressing the trehalose transporter (TRET1) and another is not. It has been thought that the intracellular water was vitrified by the intracellular trehalose, but the results of powder X-ray diffraction and differential calorimetry merely indicates the existence of ice Ih crystal below the melting point. The results of THz spectroscopy show the possibility of the presence of a liquid phase at that low temperature conditions. More detailed examinations by these measurement methods are necessary in the future.

Academic Significance and Societal Importance of the Research Achievements

「細胞の凍結保存」技術は、水産業・畜産業では既存の技術として半世紀前より利用されている。しかしそのため近年の進歩した科学技術による学術的解明は進んでおらず、凍結・保存プロトコルの改善策の検討や凍結困難細胞種への適応技術についての研究は遅れている。本研究成果は、凍結保存のプロセスを科学的に明らかにすることを目的としており、それによる既存技術の改良や、需要の高い凍結困難細胞腫(たとえばニューロンや心筋細胞等)への適応技術開発に資する。

Report

(3 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • Research Products

    (11 results)

All 2019 2018 2017

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (5 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Viabilities of long-term cryopreserved CHO-TRET1 cells with trehalose and DMSO2019

    • Author(s)
      T. Uchida, M. Furukawa, T. Kikawada, K. Yamazaki, K. Gohara
    • Journal Title

      Bulletin of Glaciological Research

      Volume: 37 Issue: 0 Pages: 1-9

    • DOI

      10.5331/bgr.18A03

    • NAID

      130007702136

    • ISSN
      1345-3807, 1884-8044
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 生物とガスハイドレート2019

    • Author(s)
      内田努
    • Journal Title

      えねるみくす

      Volume: 98 Pages: 52-55

    • NAID

      130007584355

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Temperature dependence of synchronized beating of cultured neonatal rat heart-cell networks with increasing age measured by multi-electrode arrays2018

    • Author(s)
      T. Uchida, R. Kitora, and K. Gohara
    • Journal Title

      Trends in Medicine

      Volume: 18 Issue: 4 Pages: 1-10

    • DOI

      10.15761/tim.1000145

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Intracellular trehalose via transporter TRET1 as a method to cryoprotect CHO-K1 cells2017

    • Author(s)
      Uchida Tsutomu、Furukawa Maho、Kikawada Takahiro、Yamazaki Kenji、Gohara Kazutoshi
    • Journal Title

      Cryobiology

      Volume: 77 Pages: 50-57

    • DOI

      10.1016/j.cryobiol.2017.05.008

    • NAID

      120006647014

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Xenon pressure dependence on the synchronized burst inhibition of rat cortical neuronal network cultured on multi-electrode arrays2017

    • Author(s)
      Uchida Tsutomu、Shimada Koichiro、Tanabe Ryutaro、Kubota Tatsuya、Ito Daisuke、Yamazaki Kenji、Gohara Kazutoshi
    • Journal Title

      IBRO Reports

      Volume: 3 Pages: 45-54

    • DOI

      10.1016/j.ibror.2017.09.001

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Thermodynamic study of residual water content in sediments in equilibrium with gas hydrates2017

    • Author(s)
      V. Istomin, E. Chuvilin, B. Bukhanov, T. Uchida
    • Journal Title

      Proc. 9th Int. Conf. Gas Hydrates (ICGH9), Denver, U.S.A.

      Volume: -

    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 細胞の凍結保存における凍結保護剤の役割に関する研究2019

    • Author(s)
      内田努、古川真帆、黄川田隆洋、山崎憲慈、郷原一寿
    • Organizer
      水関連研究ミニシンポジウム
    • Related Report
      2018 Annual Research Report
  • [Presentation] トレハロース輸送体を発現したCHO-K1細胞の凍結保存における保存期間依存性2018

    • Author(s)
      内田努、古川真帆、黄川田隆洋、山崎憲慈、郷原一寿
    • Organizer
      雪氷研究大会(2018・札幌)
    • Related Report
      2018 Annual Research Report
  • [Presentation] Storage-period dependence in cryopreservation of CHO-TRET1 cells with trehalose2018

    • Author(s)
      T. Uchida, M. Fufukawa, T. Kikawada, K. Yamazaki, K. Gohara
    • Organizer
      Int. Sympo. on Cryosphere and Biosphere
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] CHO細胞の凍結保存に最適な細胞内トレハロース濃度の推定2017

    • Author(s)
      内田努、古川真帆、黄川田隆洋、山崎憲慈、郷原一寿
    • Organizer
      第62回低温生物工学会年会
    • Related Report
      2017 Research-status Report
  • [Presentation] Thermodynamic study of residual water content in sediments in equilibrium with gas hydrates2017

    • Author(s)
      V. Istomin, E. Chuvilin, B. Bukhanov, T. Uchida
    • Organizer
      9th Int. Conf. Gas Hydrates (ICGH9)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research

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Published: 2017-07-21   Modified: 2020-03-30  

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