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Cell-fusion-based single-cell surgery and its application to epigenetic studies

Research Project

Project/Area Number 15H02009
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Nano/Microsystems
Research InstitutionThe University of Tokyo

Principal Investigator

WASHIZU Masao  東京大学, 大学院工学系研究科(工学部), 教授 (10201162)

Co-Investigator(Kenkyū-buntansha) オケヨ ケネディオモンディ  京都大学, ウイルス・再生医科学研究所, 講師 (10634652)
Project Period (FY) 2015-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥39,130,000 (Direct Cost: ¥30,100,000、Indirect Cost: ¥9,030,000)
Fiscal Year 2018: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
Fiscal Year 2017: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
Fiscal Year 2016: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
Fiscal Year 2015: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Keywordsマイクロ・ナノデバイス / 再生医療 / 静電気工学 / 細胞融合 / バイオナノテクノロジー / 移植・再生医療 / 静電気
Outline of Final Research Achievements

A novel method of cell fusion is developed that allows the 1:1 fusion among two types of cells with 80-90% yield. The method uses a micro-fluidic device with an array of orifice between two flow channels, and when the voltage is applied between the channels, filed constrictions are created at the orifice, by which the cells are attracted to the orifice to form cell pair. The method enables the mixing of gene / cytoplasmic factors / membrane channels between the fused cells. Using the method, a real-time observation is made on the initialization of a somatic cell fused with an ES cell. Based on the result, a microdevice is developed for the initialization utilizing iPS cell, having more availability compared with ES, where the cytoplasm of the somatic cell is replaced with that of the iPS cell through repeated fusion and separation.

Academic Significance and Societal Importance of the Research Achievements

マイクロデバイス内での細胞融合を用いて,1細胞レベルでの細胞質や細胞膜チャネルなどの交換を行う,細胞手術の手法の開発に成功した。この技術は,ガン細胞との融合により感作した免疫細胞を用いる免疫治療や,幹細胞の持つ初期化因子を用いて体細胞を初期化する技術の基礎的メカニズムを解明するとともに,それらの実用化への道を開くものと期待される。特に,本課題により行った,体細胞とES細胞との融合による初期化の1細胞レベルでの経時追跡観察や,体細胞とiPS細胞の細胞質の交換により細胞の初期化を行う技術の開発は,電界集中を利用した高収率の細胞融合の手法によって世界で初めて達成されたものである。

Report

(5 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (29 results)

All 2019 2018 2017 2016 2015 Other

All Journal Article (7 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 6 results,  Open Access: 4 results,  Acknowledgement Compliant: 3 results) Presentation (17 results) (of which Int'l Joint Research: 14 results,  Invited: 2 results) Remarks (3 results) Patent(Industrial Property Rights) (2 results) (of which Overseas: 1 results)

  • [Journal Article] A microfluidic device for isolating intact chromosomes from single mammalian cells and probing their folding stability by controlling solution conditions2018

    • Author(s)
      T. Takahashi, K. O. Okeyo, J. Ueda, K. Yamagata, M. Washizu, H. Oana
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 13684-13684

    • DOI

      10.1038/s41598-018-31975-5

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Nucleosomes of polyploid trophoblast giant cells mostly consist of histone variants and form a loose chromatin structure2018

    • Author(s)
      Koji Hayakawa, Kanae Terada; Tomohiro Takahashi, Hidehiro Oana, Masao Washizu
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1

    • DOI

      10.1038/s41598-018-23832-2

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Self-organization of human iPS cells into trophectoderm mimicking cysts induced by adhesion restriction using microstructured mesh scaffolds2018

    • Author(s)
      Kennedy O. Okeyo, Maiko Tanabe, Osamu Kurosawa, Hidehiro Oana, Masao Washizu
    • Journal Title

      Development, Growth & Differentiation

      Volume: 0 Issue: 3 Pages: 1-13

    • DOI

      10.1111/dgd.12430

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Nucleosomes exhibit non-uniform unwrapping along native chromatin fibers with increasing salt concentration as revealed by direct imaging in a microfluidic channel2018

    • Author(s)
      Hiroki Mori, Kennedy Omondi Okeyo, Masao Washizu and Hidehiro Oana
    • Journal Title

      Biotechnology Journal

      Volume: 13 Issue: 1 Pages: 1-8

    • DOI

      10.1002/biot.201700245

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Adhesion patterning by a novel air-lock technique enables localization and in-situ real-time imaging of reprogramming events in one-to-one electrofused hybrids2016

    • Author(s)
      Shota Sakamoto, Kennedy Okeyo, Satoshi Yamazaki, Osamu Kurosawa, Hidehiro Oana, Hidetoshi Kotera, and Masao Washizu
    • Journal Title

      Bio microfluidics

      Volume: 10(5) Issue: 5

    • DOI

      10.1063/1.4965422

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Minimization of cell-substrate interaction using suspended microstructured meshes initiates cell sheet formation by selfassembly organization2016

    • Author(s)
      Kennedy Omondi Okeyo, Osamu Kurosawa, Hidehiro Oana, Hidetoshi Kotera and Masao Washizu
    • Journal Title

      Biomed. Phys. Eng. Express

      Volume: 2 Issue: 6 Pages: 065019-065019

    • DOI

      10.1088/2057-1976/2/6/065019

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / 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
  • [Presentation] Nuclear transplantation between allogeneic cells achieved by fusion-based topological reconnection of the plasma membrane in a microfluidic system2018

    • Author(s)
      Masahiro Okanojo, Kennedy O. Okeyo, Hiroko Hanzawa, Osamu Kurosawa, Hidehiro Oana, Shizu Takeda, Masao Washizu
    • Organizer
      the 22th International Conference on Miniaturized System for Chemistry and Life Sciences (Micro-TAS2018)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] "reconstitution of an epithelial-endothelial bilayer by the micromesh culture technique employing an artificial matrigel basement membrane2018

    • Author(s)
      Kennedy Omondi Okeyo, Kai Yamada, Osamu Kurosawa, Hidehiro Oana, Masao Washizu
    • Organizer
      the 22th International Conference on Miniaturized System for Chemistry and Life Sciences (Micro-TAS2018)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Direct acquisition of individual target chromosomes isolated from single mammalian cells in a microfluidic channel for single cell/chromosome epigenetic analyses2018

    • Author(s)
      Hidehiro Oana, Tomohiro Takahashi, Kennedy O. Okeyo, Hiroshi Ochiai, and Masao Washizu
    • Organizer
      SELECTBIO Single Cell Analysis Summit 2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Toward fabrication of vascularized tissues using a bilayer co-culture approach achieved by a mesh culture technique2017

    • Author(s)
      Kai Yamada
    • Organizer
      Transducers 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Direct observation based analysis of compaction stability of individual chromosomes isolated from single mammalian cells2017

    • Author(s)
      Hidehiro Oana
    • Organizer
      micro-TAS 2017
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 細胞初期化に向けた電界集中型細胞融合による細胞核交換法の確立2017

    • Author(s)
      岡野定正弘
    • Organizer
      2017機械学会バイオエンジニアリング講演会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Organ-on-a-chip応用に向けたマイクロメッシュによる両面自由型細胞シートの作製2017

    • Author(s)
      山田快
    • Organizer
      2017機械学会バイオエンジニアリング講演会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Live imaging of somatic nuclear reprogramming using an electrofusion microfluidic device with air-lock patterned adhesion areas for fusant localization2016

    • Author(s)
      Shota Sakamoto, Kennedy O. Okeyo, Osamu Kurosawa, Hidehiro Oana and Masao Washizu
    • Organizer
      Micro-TAS 2016
    • Place of Presentation
      Dublin, Ireland
    • Year and Date
      2016-10-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Fabrication of freely standing oriented myotubes using a novel mesh culture method and analysis of calcium dynamics for functionality assay2016

    • Author(s)
      Kennedy O. Okeyo, M. Azeyanagi, Osamu Kurosawa
    • Organizer
      Micro-TAS 2016
    • Place of Presentation
      Dublin, Ireland
    • Year and Date
      2016-10-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Mechanical induction of trophoblast-like differentiation in human iPS cells2016

    • Author(s)
      Kennedy O. Okeyo, Osamu Kurosawa, Satoshi Yamazaki, .Hidehiro Oana, Hidetoshi Kotera, Hiromitsu Nakauchi, and Masao Washizu
    • Organizer
      Cell Symposia-10 Years of iPSCs
    • Place of Presentation
      Berkeley, US
    • Year and Date
      2016-09-25
    • Related Report
      2016 Annual Research Report
  • [Presentation] A microfluidic device for one-to-one cell electrofusion and its application toward the generation of dendritic cell for cancer immunotherapy2016

    • Author(s)
      Kennedy O. Okeyo, Yutaro Itagaki, Osamu Kurosawa, Miwako Narita, Hidehiro Oana, and Masao Washizu
    • Organizer
      MMB-2016
    • Place of Presentation
      Seoul, Korea
    • Year and Date
      2016-04-20
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Study of reprogramming using on-chip one-to-one electrofusion of somatic cells with embryonic stem cells2016

    • Author(s)
      Shota Sakamoto, Kennedy Okeyo, Osamu Kurosawa, Hidehiro Oana, Hidetoshi Kotera, Masao Washizu
    • Organizer
      IEEE-NEMS 2016
    • Place of Presentation
      Sendai, Japan (Sendai MEMS City)
    • Year and Date
      2016-04-17
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Cell Adhesion Control using Microstructured Meshes Induces Self-Assembly-Mediated Organoid Formation by Human iPS Cells2015

    • Author(s)
      Kennedy O. Okeyo, Osamu Kurosawa, Satoshi Yamazaki, Hidehiro Oana, Hidetoshi Kotera, and Masao Washizu
    • Organizer
      IEEE/ International Symposium on Micro-NanoMechatronics and Human Science
    • Place of Presentation
      Nagoya, Japan (名古屋大学)
    • Year and Date
      2015-11-23
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Microstructured Meshes as Platforms for Cell Culture and Induction of Self-Assembly-Mediated Organoid Formation2015

    • Author(s)
      Kennedy Omondi Okeyo, Osamu Kurosawa, Satoshi Yamazaki, Hidehiro Oana, Hidetoshi Kotera, and Masao Washizu
    • Organizer
      The 28th International Microprocesses and Nanotechnology Conference
    • Place of Presentation
      Toyama, Japan (ANA Crowne Plaza)
    • Year and Date
      2015-11-10
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Cell adhesion control by a novel mesh culture method directs differentiation and self-assembly of human iPS cells into trophoplasts; towards organ-n-chip embryogenesis2015

    • Author(s)
      Kennedy Omondi Okeyo, Osamu Kurosawa, Satoshi Yamazaki, Hidehiro Oana, Hidetoshi Kotera, and Masao Washizu
    • Organizer
      micro-TAS 2015
    • Place of Presentation
      Gyeongju, Korea
    • Year and Date
      2015-10-25
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Engineering Free Standing 2D and 3D Biostructures by Induction of Self-Assembly Organization of Cells in a Suspension2015

    • Author(s)
      Kennedy Okeyo and Masao Washizu
    • Organizer
      Lab-on-a-Chip, Microfluidics & Microarrays World Congress
    • Place of Presentation
      San Diego, USA
    • Year and Date
      2015-09-28
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] On-chip high-yield electrofusion by electric field constriction and culture of fusants on air-lock patterned adhesion areas2015

    • Author(s)
      Shota Sakamoto, Kennedy O. Okeyo, Osamu Kurosawa, Hidehiro Oana, Hidetoshi Kotera and Masao Washizu
    • Organizer
      7th International Symposium on Microchemistry and Microsystems 2015
    • Place of Presentation
      Kyoto, Japan (京都大学)
    • Year and Date
      2015-06-08
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Remarks] 東京大学大学院工学系研究科鷲津・小穴研究室

    • URL

      http://www.washizu.t.u-tokyo.ac.jp/

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
  • [Remarks] www.washizu.t.u-tokyo.ac.jp

    • Related Report
      2016 Annual Research Report
  • [Remarks] www.washizu.t.u-tokyo.ac.jp

    • Related Report
      2015 Annual Research Report
  • [Patent(Industrial Property Rights)] 細胞製造装置2019

    • Inventor(s)
      岡野定雅弘・武田志津・オケヨケネディオモンディ・鷲津正夫
    • Industrial Property Rights Holder
      株式会社日立製作所・国立大学法人東京大学
    • Industrial Property Rights Type
      特許
    • Filing Date
      2019
    • Related Report
      2018 Annual Research Report
    • Overseas
  • [Patent(Industrial Property Rights)] 細胞質の交換方法、細胞の製造方法、及び装置2016

    • Inventor(s)
      岡野定雅弘・武田志津・オケヨケネディオモンディ・鷲津正夫
    • Industrial Property Rights Holder
      岡野定雅弘・武田志津・オケヨケネディオモンディ・鷲津正夫
    • Industrial Property Rights Type
      特許
    • Filing Date
      2016-10-10
    • Related Report
      2016 Annual Research Report

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Published: 2015-04-16   Modified: 2020-03-30  

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