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A study of transporting molecules into cell

Research Project

Project/Area Number 23246045
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Intelligent mechanics/Mechanical systems
Research InstitutionKyoto University

Principal Investigator

Kotera Hideyoshi  京都大学, 工学(系)研究科(研究院), 教授 (20252471)

Co-Investigator(Kenkyū-buntansha) WASHIZU Masao  東京大学, 大学院工学系研究科, 教授 (10201162)
Project Period (FY) 2011-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥48,230,000 (Direct Cost: ¥37,100,000、Indirect Cost: ¥11,130,000)
Fiscal Year 2015: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2014: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
Fiscal Year 2013: ¥10,010,000 (Direct Cost: ¥7,700,000、Indirect Cost: ¥2,310,000)
Fiscal Year 2012: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
Fiscal Year 2011: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
Keywordsナノ・マイクロ科学 / マイクロ・ナノデバイス / 微細加工 / 細胞融合 / 初期化 / 再生医療 / 静電気 / iPS細胞 / Micro TAS / electroporation / ヒトiPS / 力場 / マイクロ構造 / 機能発現 / マイクロデバイス / 細胞機能 / 細胞の初期化 / ヒトiPS細胞 / 弱い力場 / エレクトロポーレーション / 細胞環境 / マイクロ流体デバイス / ナノ・マイクロ電気穿孔 / 遺伝子導入 / 物質輸送 / マイクロファブリケーション / 電気泳動 / 拡散 / マイクロ・ナノメカトロニクス / マイクロTAS / iPS / 細胞 / エレクトロポレーション
Outline of Final Research Achievements

We have developed a novel gene delivery system based on on-chip electroporation, where plasmids are directly transported into the nucleus by electrophoresis. The basic concept is examined by micro orifice fabricated on vertical PDMS wall with quantum dot and FEM simulation. The cells are cultured on micro-orifices and placed between electrodes, to which a electric pulse was applied in a DMEM medium containing GFP plasmids. Most cells showed GFP expression after pulsation, suggesting that the plasmid was directly fed into the cell nucleus. Then we applied the system to produce iPS cells. After introduction of Yamanaka factors, cells were cultured on the orifice for three weeks after electroporation, and then reprogramming of nucleus was checked. The fact that the plasmids are instantaneously transported directly into nucleus of selected cells, with precisely controllable timing, suggests that the method will find its applications in re-programming or differentiation control studies.

Report

(6 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • Research Products

    (27 results)

All 2015 2014 2013 2012 Other

All Journal Article (6 results) (of which Peer Reviewed: 6 results,  Acknowledgement Compliant: 4 results,  Open Access: 3 results) Presentation (18 results) (of which Int'l Joint Research: 5 results,  Invited: 4 results) Remarks (3 results)

  • [Journal Article] Characterization of optically-driven microstructures for manipulating single DNA molecules under a fluorescence microscope2015

    • Author(s)
      K. Terao*, C. Masuda, R. Inukai, M. Gel, H. Oana, M. Washizu, T. Suzuki, H. Takao, F. Shimokawa, F. Oohira
    • Journal Title

      IET Nanobiotechnology

      Volume: - Issue: 3 Pages: 10-10

    • DOI

      10.1049/iet-nbt.2015.0036

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Cell Adhesion Minimization by a Novel Mesh Culture Method Mechanically Directs Trophoblast Differentiation and Self-Assembly Organization of Human Pluripotent Stem Cells2015

    • Author(s)
      Okeyo O. Kennedy, Osamu Kurosawa, Satoshi Yamazaki, Hidehiro Oana, Hidetoshi Kotera, Hiromitsu Nakauchi, Masao Washizu
    • Journal Title

      Tissue Engineering Part C

      Volume: なし Issue: 10 Pages: 1105-15

    • DOI

      10.1089/ten.tec.2015.0038

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Development of low-fluorescence thick photoresist for high-aspect-ratio microstructure in bio-application2015

    • Author(s)
      Tamai, H. Maruo, K. Ueno, H. Terao, K. Kotera, H. Suzuki, T.
    • Journal Title

      Biomicrofluidic

      Volume: 9

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Single-cell cloning and expansion of human induced pluripotent stem cells by a microfluidic culture device2014

    • Author(s)
      Taku Matsumura, Kazuya Tatsumi, Yuichiro Noda, Naoyuki Nakanishi, Atsuhito Okonogi, Kunio Hirano, Liu Li, Takashi Osumi, Takashi Tada, Hidetoshi Kotera
    • Journal Title

      Biochemical and Biophysical Research Communications

      Volume: 453 Issue: 1 Pages: 131-137

    • DOI

      10.1016/j.bbrc.2014.09.081

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] オンチップエレクトロポレーションを用いた接着細胞核への遺伝子直 接送達法の開発2014

    • Author(s)
      黒澤修・オケヨ ケネディ オモンディ・小穴英廣・沖田圭介・小寺秀俊・鷲 津正夫
    • Journal Title

      静電気学会誌

      Volume: Vol.38, No.1 Pages: 28-33

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] ubcellular glucose exposure biases the spatial distribution of insulin granules in single pancreatic beta cells2014

    • Author(s)
      Kyohei Terao, Murat Gel, Atsuhito Okonogi, Ariko Fuke, Teru Okitsu, Takashi Tada, Takaaki Suzuki, Shinya Nagamatsu, Masao Washizu & Hidetoshi Kotera
    • Journal Title

      Scientific Reports

      Volume: 4 Issue: 1 Pages: 1-6

    • DOI

      10.1038/srep04123

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Mapping positional distribution of higher-order structures along unfragmented native chromatin fibers isolated from single cells in a microchannel2015

    • Author(s)
      Hiroki Mori, Kennedy Omondi Okeyo, Masao Washizu and Hidehiro Oana
    • Organizer
      micro-TAS 2015
    • Place of Presentation
      Gyeongju, Kore
    • Year and Date
      2015-10-25
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Developing single cell culture platform for human pluripotent stem cells2015

    • Author(s)
      Lenin yu
    • Organizer
      JCK MEMS 2015 Conference
    • Place of Presentation
      Xi’an China
    • Year and Date
      2015-09-23
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Importance of substrate and interaction between cell for culturing and realizing nano-human2015

    • Author(s)
      Leqian Yu, Li Liu, Junjun Li, Yong Chen, Hitrohumi Shintaku, Ryuji Yokokawa, Takaaki Suzuki, Masao Washizu, Kenedy M. Okeyo and Hidetoshi Kotera
    • Organizer
      2nd Africa International Biotechnology and Biomedical Conference (AIBBC 2015)
    • Place of Presentation
      Nirobi
    • Year and Date
      2015-09-17
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] SINGLE CELL CULTURE OF HUMAN PLURIPOTENT STEM CELLS2015

    • Author(s)
      Leqian Yu, Li Liu, Junjun Li, Wei Yang, Fuchou Tang, Nanae Fujimoto, Minako Nakajima, Yong Chen1, Hidetoshi Kotera
    • Organizer
      International Society for Stem Cell Research 2015
    • Place of Presentation
      Stockholm, Sweden
    • Year and Date
      2015-06-24
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Technique for Cell Adhesion Control and Initiation of Cellular Structure Formation Employing Suspended Microstructured Meshes2015

    • Author(s)
      Kennedy O. Okeyo, Rina Yanaru, Osamu Kurosawa, Satoshi Yamazaki, Hidehiro Oana, Hidetoshi Kotera, Masao Washizu
    • Organizer
      ISMM 7th International Symposium on Microchemistry and Microsystems
    • Place of Presentation
      Kyoto,Japan
    • Year and Date
      2015-06-08
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Interactive Bio Surface of Micro Fluidic Device for Organ on a Chip2015

    • Author(s)
      H.Kotera
    • Organizer
      The 6th Taiwan- Japan Symposium on Nanomedicene
    • Place of Presentation
      Taiwan Academia Sinica
    • Year and Date
      2015-01-08 – 2015-01-09
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] CYTOPLASMIC TRANSFER WITHOUT NUCLEI MIXING BETWEEN DENDRITIC CELLS AND TUMOR CELLS ACHIEVED BY ONE-TO-ONE ELECTROFUSION VIA MICRO-ORIFICES IN A MICROFLUIDIC DEVICE2014

    • Author(s)
      Yutaro Itagaki , Kennedy O. Okeyo, Osamu Kurosawa, Hidehiro Oana, Miwako Narita, Hidetoshi Kotera, and Masao Washizu
    • Organizer
      The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      San Antonio, TX, USA
    • Year and Date
      2014-10-26 – 2014-10-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] STABLE AND LONG TERM CULTURE OF STEM CELLS UNDER SHEAR FLOW ON A MICROSTRUCTURED MESH SHEET EMBEDDED IN A FLUIDIC CHAMBER2014

    • Author(s)
      K.O. Okeyo, T. Isozaki, O. Kurosawa, H. Oana, H. Kotera and M. Washizu
    • Organizer
      The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      San Antonio, TX, USA
    • Year and Date
      2014-10-26 – 2014-10-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] GENERATION OF EPITHELIAL CELL SHEETS WITH DEFINED CELL ORIENTATION USING MICROSTRUCTURED MESH SHEETS AS SUBSTRATES FOR CELL CULTURE2014

    • Author(s)
      K. O. Okeyo, R. Yanaru, O. Kurosawa, H. Oana, H. Kotera and M. Washizu
    • Organizer
      The 18th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      San Antonio, TX, USA
    • Year and Date
      2014-10-26 – 2014-10-30
    • Related Report
      2014 Annual Research Report
  • [Presentation] Function of single cell and tissue for regenerative medicine2014

    • Author(s)
      H.Kotera
    • Organizer
      Joint Meeting of the 1st Africa International Biotechnology & Biomedical Conference and the 8th International Workshop on Approaches to Single-Cell Analysis,
    • Place of Presentation
      Nairobi, Kenya
    • Year and Date
      2014-09-10 – 2014-09-12
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 分子濃縮によるオンチップ電気穿孔法の効率化2013

    • Author(s)
      重里 優子, 横川 隆司, 小寺 秀俊, 新宅 博文
    • Organizer
      第5回マイクロ・ナノ工学シンポジウム
    • Place of Presentation
      仙台
    • Year and Date
      2013-11-05 – 2013-11-07
    • Related Report
      2013 Annual Research Report
  • [Presentation] On-chip electroporation device for direct introduction of plasmids into cell nucleus and observation of cell reprogramming process2013

    • Author(s)
      K. O. Okeyo, Y. Hayashi, O. Kurosawa, H. Oana, H. Kotera, M. Washizu
    • Organizer
      µ-TAS 2013
    • Place of Presentation
      Freiburg, Germany
    • Year and Date
      2013-10-27 – 2013-10-31
    • Related Report
      2013 Annual Research Report
  • [Presentation] Cell adhesion control initiate cell sheet formation in a medium suspension2013

    • Author(s)
      K. O. Okeyo, N. Omasa, O. Kurosawa, H. Oana, H. Kotera, M. Washizu
    • Organizer
      µ-TAS 2013
    • Place of Presentation
      Freiburg, Germany
    • Year and Date
      2013-10-27 – 2013-10-31
    • Related Report
      2013 Annual Research Report
  • [Presentation] オンチップエレクトロポレーションを用いた接着細胞核への遺伝子直接送達法の開発2013

    • Author(s)
      黒澤修・オケヨ ケネディ オモンディ・小穴英廣・沖田圭介・小寺秀俊・鷲津正夫
    • Organizer
      静電気学会第37回全国大会
    • Place of Presentation
      千葉大学
    • Year and Date
      2013-09-10 – 2013-09-11
    • Related Report
      2013 Annual Research Report
  • [Presentation] 電界集中型細胞融合法による抗体産生細胞の高収率取得2013

    • Author(s)
      オケヨ ケネディ・西垣内康宏・近藤武宏・黒澤修・小穴英廣・小寺秀俊・鷲津正夫
    • Organizer
      静電気学会第37回全国大会
    • Place of Presentation
      千葉大学
    • Year and Date
      2013-09-10 – 2013-09-11
    • Related Report
      2013 Annual Research Report
  • [Presentation] エレクトロポレーションにおける導入物質観察の為の微小流路デバイス2012

    • Author(s)
      梶本剛生 小此木孝仁 鈴木孝明 新宅 博文 横川隆司 小寺秀俊
    • Organizer
      日本機械学会 2012年度年次大会
    • Place of Presentation
      金沢
    • Related Report
      2012 Annual Research Report
  • [Presentation] 光応答性タンパク質による細胞刺激のためのマイクロデバイス2012

    • Author(s)
      重里優子 駒井章治 小此木孝仁 大岡正孝 巽和也 新宅博文 横川 隆司 小寺 秀俊
    • Organizer
      日本機械学会 2012年度年次大会
    • Place of Presentation
      金沢
    • Related Report
      2012 Annual Research Report
  • [Presentation] 電界集中を用いた細胞の電気穿孔法における物質輸送の可視化計測2012

    • Author(s)
      梶本剛生 新宅 博文 横川 隆司 小寺 秀俊
    • Organizer
      第4回マイクロ・ナノ工学シンポジウム
    • Place of Presentation
      小倉
    • Related Report
      2012 Annual Research Report
  • [Remarks] マイクロエンジニアリング専攻 ナノシステム創成工学講座 ナノメトリックス工学分野

    • URL

      http://www.ksys.me.kyoto-u.ac.jp

    • Related Report
      2015 Annual Research Report
  • [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/xT6yD

    • Related Report
      2014 Annual Research Report 2013 Annual Research Report
  • [Remarks] researchmap

    • URL

      http://researchmap.jp/read0042655

    • Related Report
      2014 Annual Research Report

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Published: 2011-06-18   Modified: 2019-07-29  

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