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Standardization of embryoid body formation from embryonic stem cellsand quality estimation of embryoid bodies

Research Project

Project/Area Number 18560751
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionUniversity of Yamanashi

Principal Investigator

KUROSAWA Hiroshi  University of Yamanashi, Department of Research Interdisciplinary Graduate School of Medicine and Engineering, Associate Professor (10225295)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,580,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsES cell / embryoid body / quantitative PCR / EB formation method / deifferentiation / gene expression / standardization / RT-PCR / 発現遺伝子
Research Abstract

The gene expression of EBs was quantitatively analyzed by real-time PCR and the EBs were standardized based on the data of real-time PCR.
1. Formation of various types of EBs Various types of EBs were famed in different culture conditions of culture vessel, seeding cell density, oxygen, and nutrient. The EBs were famed by hanging drop method or in low-adherence 96-well multi-plate.
2. Gene analysis of EBs by real-time PCR
Marker genes (Oct3/4, Rex1, GATA4, GATA6, TTR, AFP, α-MHC, Nkx2.5), which represent differentiation status of EBs, were quantitatively analyzed by real-time PCR.
3. Graphical presentation of gene expression pattern of EBs The gene expression pattern of EBs was graphically shown to characterize the differentiation status of EBs.
4. Making sure of the relationship between gene expression pattern and actual differentiation observed in the EB differentiation culture
The EBs highly expressed of a-MHC and Nkx2.5 generate cardiomyocytes with high efficiency. The EBs highly expressed of FIR and AFP have a trend to generate visceral yolk-sac-like structure. It was found that the gene egression pattern in EBs directly reflected the differentiation tendency to be observed in the following culture of EBs.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (13 results)

All 2007 Other

All Journal Article (5 results) (of which Peer Reviewed: 3 results) Presentation (6 results) Remarks (2 results)

  • [Journal Article] Methods for inducing embryoid body formation: In vitro differentiation system of embryonic stem cells.2007

    • Author(s)
      Hiroshi Kurosawa
    • Journal Title

      Journal of Bioscience and Bioengineering 103

      Pages: 389-398

    • NAID

      110006278720

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies.2007

    • Author(s)
      Mikiko Koike
    • Journal Title

      Journal of Bioscience and Bioengineering 104

      Pages: 294-299

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Methods for inducing embryoid body formation : In vitro differentiation system of embryonic stem cells2007

    • Author(s)
      Hiroshi, Kurosawa
    • Journal Title

      Journal of Bioscience and Bioengineering Vol.103, No.5

      Pages: 389-398

    • NAID

      110006278720

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies2007

    • Author(s)
      Mikiko, Koike, Shujiro, Sakaki, Yoshifumi, Amano, Hiroshi Kurosawa
    • Journal Title

      Journal of Bioscience and Bioengineering Vol.104, No.4

      Pages: 294-299

    • NAID

      110006437010

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Characterization of embryoid bodies of mouse embryonic stem cells formedunder various culture conditions and estimation of differentiation status ofsuch bodies.2007

    • Author(s)
      Mikiko Koike
    • Journal Title

      Journal of Bioscience and Bioengineering 104

      Pages: 294-299

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] Graphical presentation of gene expression pattern in various types of embryoid bodies from mouse ES cells.2007

    • Author(s)
      Mikiko Koike
    • Organizer
      TERMIS-AP 2007
    • Place of Presentation
      グランドプリンスホテル赤坂
    • Year and Date
      2007-12-04
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] Graphical presentation of gene expression pattern in various types of embryoid bodies from mouse ES cells2007

    • Author(s)
      Mikiko, Koike, Shujiro, Sakaki, Yoshifumi, Amano, Hiroshi, Kurosawa
    • Organizer
      TERMIS AP 2007
    • Place of Presentation
      Tokyo
    • Year and Date
      2007-12-04
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 各種マウスES細胞胚様体における遺伝子発現パターンの図式化2007

    • Author(s)
      小池 美紀子
    • Organizer
      第59回日本生物工学会大会
    • Place of Presentation
      広島大学
    • Year and Date
      2007-09-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] マウスES細胞由来嚢胞性胚様体における形成条件の最適化と分化傾向の特徴2007

    • Author(s)
      安田 恵美子
    • Organizer
      第59回日本生物工学会大会
    • Place of Presentation
      広島大学
    • Year and Date
      2007-09-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] Graphical presentation of gene expression pattern in various types of embryoid bodies from mouse ES cells2007

    • Author(s)
      Mikiko, Koike, Shujiro, Sakaki, Yoshifumi, Amano, Hiroshi, Kurosawa
    • Organizer
      Annual Meeting 2007 The Society for Biotechnology (Japan)
    • Place of Presentation
      Hiroshima University, Hiroshima
    • Year and Date
      2007-09-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Optimization of cystic embryoid body (EB) formation from mouse ES cells and expression of marker genes in differentiating EBs2007

    • Author(s)
      Emiko, Yasuda, Yuji, Seki, Shuiiro, Sakaki. Hiroshi, Kurosawa
    • Organizer
      Annual Meeting 2007 The Society for Biotechnology(Japan)
    • Place of Presentation
      Hiroshima University. Hiroshima
    • Year and Date
      2007-09-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Remarks] 「研究成果報告書概要(和文)」より

    • URL

      http://erdb.yamanashi.ac.jp/rdb/A_DispInfo.Scholar?ID=BC049428387331DC

    • Related Report
      2007 Final Research Report Summary
  • [Remarks]

    • URL

      http://erdb.yamanashi.ac.jp/rdb/A_DispInfo.Scholar?ID=BC049428387331DC

    • Related Report
      2007 Annual Research Report

URL: 

Published: 2006-04-01   Modified: 2016-04-21  

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