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Safe and efficient selection of differentiated cells by stage-specific gene expression from auto-erasable vector

Research Project

Project/Area Number 19K22945
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 90:Biomedical engineering and related fields
Research InstitutionUniversity of Tsukuba

Principal Investigator

Nishimura Ken  筑波大学, 医学医療系, 准教授 (80500610)

Project Period (FY) 2019-06-28 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2019: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywords分化誘導 / 再生医療 / 創薬 / 自動除去型SeVdpベクター / miRNA / 神経幹細胞 / SeVdpベクター / 分化細胞 / 分化促進遺伝子
Outline of Research at the Start

本研究では、高純度分化細胞の単離のために、分化誘導効率の向上と分化細胞選択の最適化の両方を、独自の自動除去型SeVdpベクターを用いて実現させる。誘導効率向上のためには、各分化段階特異的なmiRNAを利用して、ベクターからの遺伝子発現を分化段階に合わせてコントロールする。また分化細胞選択のために、目的分化細胞特異的に発現するmiRNAによって分化細胞からのみベクターが自動除去されるようにして、分化細胞のみを選択する。そして最終的に、ベクターを一度導入するのみで高効率高純度分化誘導を実現させる。

Outline of Final Research Achievements

To enhance an application of differentiated tissue to regenerative medicine and drug discovery, we need to reduce risks of tumor formation by remained undifferentiated cells and obtain highly pure differentiated cells efficiently. Adding target sequences of neural stem cell (NSC)-specific miRNA to our original auto-erasable SeVdp vector enabled us to select pure population of the induced NSCs. Moreover, we succeeded to control an expression timing of a NSC differentiation inducer from SeVdp vector by miRNA whose expression is changed at a particular period in the NSC differentiation. We plan to combine the cell selection system and the differentiation induction into single SeVdp vector to establish a system for efficient selection of highly pure differentiated cells.

Academic Significance and Societal Importance of the Research Achievements

本研究成果によって、再生医療や創薬に実用化可能な分化細胞を取得するために必要な技術の開発が進展した。この技術を完成させるためには、ベクターコピー数の微調整や、ベクター除去に用いるために最適なmiRNAの探索、試行などが必要ではあるが、そのような研究を継続して高効率高純度分化細胞選択系を確立することによって、幹細胞等から誘導して作製した分化細胞を用いた、安全な再生医療や新しい医薬の探索が可能になることが期待される。

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (6 results)

All 2021 2020 2019

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results,  Open Access: 3 results) Presentation (2 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Journal Article] Efficient Reprogramming of Canine Peripheral Blood Mononuclear Cells into Induced Pluripotent Stem Cells2021

    • Author(s)
      Kimura Kazuto、Tsukamoto Masaya、Tanaka Miyuu、Kuwamura Mitsuru、Ohtaka Manami、Nishimura Ken、Nakanishi Mahito、Sugiura Kikuya、Hatoya Shingo
    • Journal Title

      Stem Cells and Development

      Volume: 30 Issue: 2 Pages: 79-90

    • DOI

      10.1089/scd.2020.0084

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Functional analysis of PTH1R variants found in primary failure of eruption.2020

    • Author(s)
      Izumida E, Suzawa T, Miyamoto Y, Yamada A, Takimoto R, Otsu M, Saito T, Yamaguchi T, Nishimura K, Ohtaka M, Nakanishi M, Yoshimura K, Sasa K, Takimoto R, Uyama R, Shirota T, Maki K, Kamijo R.
    • Journal Title

      Journal of Dental Research

      Volume: 99 Issue: 4 Pages: 429-436

    • DOI

      10.1177/0022034520901731

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] 4-Phenylbutyrate ameliorates apoptotic neural cell death in Down syndrome by reducing protein aggregates2020

    • Author(s)
      Hirata Katsuya、Nambara Toshihiko、Kawatani Keiji、Nawa Nobutoshi、Yoshimatsu Hidetaka、Kusakabe Haruna、Banno Kimihiko、Nishimura Ken、Ohtaka Manami、Nakanishi Mahito、Taniguchi Hidetoshi、Arahori Hitomi、Wada Kazuko、Ozono Keiichi、Kitabatake Yasuji
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 14047-14047

    • DOI

      10.1038/s41598-020-70362-x

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Induction of Noonan syndrome-specific human-induced pluripotent stem cells under serum-, feeder-, and integration-free conditions2020

    • Author(s)
      Atsuko Hamada, Eri Akagi, Fumitaka Obayashi, Sachiko Yamasaki, Koichi Koizumi, Manami Ohtaka, Ken Nishimura, Mahito Nakanishi, Shigeaki Toratani & Tetsuji Okamoto
    • Journal Title

      In Vitro Cellular & Developmental Biology - Animal

      Volume: 56(10) Issue: 10 Pages: 888-895

    • DOI

      10.1007/s11626-020-00515-9

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] 再生医療や創薬研究を支援する高純度分化細胞選択法2020

    • Author(s)
      西村健
    • Organizer
      BioJapan2020
    • Related Report
      2020 Annual Research Report
  • [Presentation] 安全な再生医療を実現させる高純度分化細胞選択法2019

    • Author(s)
      西村健
    • Organizer
      日中大学フェア&フォーラム in CHINA 2019
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research / Invited

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Published: 2019-07-04   Modified: 2024-12-25  

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