Basic research on the creation of innovative human iPS cell culture processes using complex physical stimulation.

• Project/Area Number: 18K14063
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: Kyoto University
• Principal Investigator: Horie Masanobu 京都大学, 環境安全保健機構, 助教 (60727014)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: メカノバイオロジー / ヒトiPS細胞 / ティッシュエンジニアリング / 自己組織化 / 骨格筋 / 培養プロセス / 生物化学工学 / 大量培養 / ヒト多能性幹細胞細胞 / 細胞製造 / 物理刺激 / ヒト幹細胞 / 軟培養面 / 電場 / バイオリアクター / メカノトランスダクション / デバイス

Outline of Final Research Achievements

We have succeeded in creating an inexpensive soft culture surface for controlling the behavior of human iPS cells. It was clarified that undifferentiated maintenance culture of human iPS cells can be performed only by controlling the hardness of the culture surface without adding expensive cytokines used for maintenance of undifferentiation.
It was clarified that the self-organization of mouse skeletal myoblasts is induced depending on the hardness of the culture surface and ECM. Furthermore, we succeeded in constructing contracting artificial skeletal muscle cells without using scaffolding such as gel or scaffold.

Academic Significance and Societal Importance of the Research Achievements

物理刺激のみによってヒトiPS細胞の未分化維持培養ができれば、現在高コストが実用化障壁の1つになっているヒトiPS細胞を用いた細胞治療のための、新たな培養プロセス開発の一助となる。
また、自己組織化を効率的に誘導するプロセスを開発したことによって、より生体内の状態に近い人工骨格筋組織を誘導することができ、世界的な動物実験に対する風当たりの中において、新たな創薬等の研究開発のためのツールとなり得ると考えられる。

Research Products

• [Journal Article] A Novel Method for Polyacrylamide Gel Preparation Using N-hydroxysuccinimide-acrylamide Ester to Study Cell-Extracellular Matrix Mechanical Interactions (2021)
  • Author(s): Kumai Jun、Sasagawa Satoru、Horie Masanobu、Yui Yoshihiro
  • Journal Title: Frontiers in Materials Volume: 8 Pages: 637278-637278
  • DOI: 10.3389/fmats.2021.637278
  • Peer Reviewed / Open Access
• [Journal Article] Effect of hydrogen peroxide concentration on the maintenance and differentiation of cultured skeletal muscle cells. (2021)
  • Author(s): Hideaki Fujita, Keisuke Mae, Kouki Nagatani, Masanobu Horie, Eiji Nagamori
  • Journal Title: Journal of Bioscience and Bioengineering Volume: 131 Issue: 5 Pages: 572-578
  • DOI: 10.1016/j.jbiosc.2020.12.010
  • NAID: 40022568974
  • Peer Reviewed
• [Journal Article] The effect of cell-extracellular matrix interaction on myogenic characteristics and artificial skeletal muscle tissue. (2020)
  • Author(s): Masanobu Horie, Ran Ding, Sumire Nagasaka, Saki Ohsumi, Kazunori Shimizu, Hiroyuki Honda, Eiji NAGAMORI, Hideaki Fujita, Takuo Kawamoto
  • Journal Title: Journal of Bioscience and Bioengineering Volume: in press Issue: 1 Pages: 98-105
  • DOI: 10.1016/j.jbiosc.2020.02.008
  • NAID: 40022293633
  • Peer Reviewed
• [Journal Article] The effect of cell-extracellular matrix interaction on myogenic characteristics and artificial skeletal muscle tissue (2020)
  • Author(s): Ran Ding, Masanobu Horie, Sumire Nagasaka, Saki Ohsumi, Kazunori Shimizu, Hiroyuki Honda, Eiji Nagamori, Hideaki Fujita, Takuo Kawamoto
  • Journal Title: Journal of Bioscience and Bioengineering Volume: In press
  • NAID: 40022293633
  • Peer Reviewed
• [Journal Article] Linking substrate and nucleus via actin cytoskeleton in pluripotency maintenance of mouse embryonic stem cells (2019)
  • Author(s): David Brit Gracy、Fujita Hideaki、Yasuda Kyota、Okamoto Kazuko、Panina Yulia、Ichinose Junya、Sato Osamu、Horie Masanobu、Ichimura Taro、Okada Yasushi、Watanabe Tomonobu M
  • Journal Title: Stem Cell Research Volume: 41 Pages: 101614-101614
  • DOI: 10.1016/j.scr.2019.101614
  • Peer Reviewed / Open Access
• [Journal Article] Targeted suppression of metastasis regulatory transcription factor SOX2 in various cancer cell lines using a sequence-specific designer pyrrole-imidazole polyamide (2019)
  • Author(s): Madhu Malinee, Alok Kumar, Takuya Hidaka, Masanobu Horie, Kouichi Hasegawa, Ganesh N Pandian, Hiroshi Sugiyama
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 28 Issue: 3 Pages: 115248-115248
  • DOI: 10.1016/j.bmc.2019.115248
  • Peer Reviewed / Open Access
• [Journal Article] Inkjet micropatterning through horseradish peroxidase-mediated hydrogelation for controlled cell immobilization and microtissue fabrication (2019)
  • Author(s): Gantumur, Enkhtuul, Kimura, Miyu, Taya, Masahito, Horie, Masanobu, Nakamura, Makoto, Sakai, Shinji
  • Journal Title: Biofacrication Volume: 12 Issue: 1 Pages: 011001-011001
  • DOI: 10.1088/1758-5090/ab3b3c
  • Peer Reviewed / Open Access
• [Presentation] SOFT SUBSTRATES PROMOTE SELF-RENEWAL OF HUMAN INDUCED PLURIPOTENT STEM CELLS IN BFGF REDUCED CONDITION (2020)
  • Author(s): Masanobu Horie
  • Organizer: International Society for Stem Cell Research Annual Meeting 2020
  • Int'l Joint Research
• [Presentation] Substrate stiffness dependent growth properties of human iPS cells (2019)
  • Author(s): Masanobu Horie
  • Organizer: Termi-AP 2019
  • Int'l Joint Research
• [Presentation] 高効率ヒトiPS細胞由来骨格筋細胞誘導および組織構築プロセスの開発 (2019)
  • Author(s): Masanobu Horie
  • Organizer: JAACT2019
• [Presentation] Self-organization of mouse myoblast cells mediated by soft substrates and improvement of myotube formations in artificial skeletal muscle (2019)
  • Author(s): Ran Ding
  • Organizer: Termi-AP 2019
  • Int'l Joint Research
• [Presentation] ヒトiPS細胞の筋分化誘導条件の検討 (2019)
  • Author(s): 吉開太一、井藤彰、池田一史、吉岡貫太郎、堀江 正信、櫻井英俊、堀田秋津、河邉佳典、上平正道
  • Organizer: 化学工学会第84年会
• [Presentation] 複数素材が適用可能なインクジェット式バイオプリンターを利用した細胞パターニングの構築 (2018)
  • Author(s): Gantumur Enkhtuul、木村美夢、堀江正信、境慎司、中畑雅樹、田谷正仁
  • Organizer: 化学工学会第50回秋季大会
• [Presentation] ヒトiPS細胞由来筋芽細胞の高機能分化誘導法の開発 (2018)
  • Author(s): 吉開太一、井藤彰、池田一史、吉岡貫太郎、堀江正信、河邉佳典、上平正道
  • Organizer: 化学工学会第50回秋季大会
• [Presentation] 自己組織化現象を用いた人工骨格筋組織の構築 (2018)
  • Author(s): 丁然、堀江正信、川本卓男
  • Organizer: 化学工学会第50回秋季大会
• [Presentation] Substrate stiffness dependent growth properties of human induced pluripotent stem cells (2018)
  • Author(s): Masanobu Horie
  • Organizer: Biotechnology Towards Next generation single Cell Analysis
  • Int'l Joint Research / Invited
• [Book] 骨格筋社会を核とした筋スマート社会 (2019)
  • Author(s): 堀江正信、櫻井英俊
  • Publisher: シーエムシー
• [Remarks] 京都大学教育研究活動データベース
  • URL: https://kyouindb.iimc.kyoto-u.ac.jp/j/oL4aF