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In vitro method for detection of immortalized cells in human retinal pigment epithelial cells.

Research Project

Project/Area Number 25871244
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Drug development chemistry
Ophthalmology
Research InstitutionNational Institute of Health Sciences (2014-2015)
Foundation for Biomedical Research and Innovation (2013)

Principal Investigator

Kuroda Takuya  国立医薬品食品衛生研究所, 再生・細胞医療製品部, 研究員 (70648857)

Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywords不死化細胞 / 網膜色素上皮細胞 / 造腫瘍性 / iPS細胞 / 造腫瘍性細胞
Outline of Final Research Achievements

Human pluripotent stem cells (hPSCs), i.e. human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are able to self-renew and differentiate into multiple cell types. Thus, numerous attempts have been made to utilize hPSCs in regenerative medicine/cell therapy. However hPSCs have tumorigenic potential. Therefore, residual undifferentiated hPSCs or contamination of the transformed cells in products that would eventually proliferate and form a teratoma is one of the most obvious safety issues to develop cell therapy hPSC-derived products. In the present study, we identified a new immortalized RPE cell marker and developed new qRT-PCR method to detect contamination of human RPE cells with immortalized RPE cells. This qRT-PCR assay can detect as few as 3% of immortalized RPE cells in normal RPE cells. This in vitro qRT-PCR method is expected to contribute to process validation and quality control of cell therapy products derived from hPSCs.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (14 results)

All 2016 2015 2014

All Journal Article (4 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (9 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results) Book (1 results)

  • [Journal Article] Highly sensitive droplet digital PCR method for detection of residual undifferentiated cells in cardiomyocytes derived from human pluripotent stem cells.2015

    • Author(s)
      Kuroda T, Yasuda S, Matsuyama S, Tano K, Kusakawa S, Sawa Y, Kawamata S, Sato Y.
    • Journal Title

      Regen. Ther.

      Volume: 2 Pages: 17-23

    • DOI

      10.1016/j.reth.2015.08.001

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Ultra-sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products by digital analysis of soft agar colony formation.2015

    • Author(s)
      Kusakawa S, Yasuda S, Kuroda T, Kawamata S, Sato Y.
    • Journal Title

      Sci. Rep.

      Volume: 5 Issue: 1 Pages: 17892-17892

    • DOI

      10.1038/srep17892

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Characterization of in vivo tumorigenicity tests using severe immunodeficient NOD/Shi-scid IL2Rgnull mice for detection of tumorigenic cellular impurities in human cell-processed therapeutic products.2015

    • Author(s)
      Kusakawa S, Machida K, Yasuda S, Takada N,; Kuroda T, Sawada R, Okura H, Tsutsumi H, Kawamata S, Sato Y
    • Journal Title

      REGENERATIVE THERAPY

      Volume: Volume 1 Pages: 30-37

    • DOI

      10.1016/j.reth.2014.12.001

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A novel in vitro method for detecting undifferentiated human pluripotent stem cells as impurities in cell therapy products using a highly efficient culture system.2014

    • Author(s)
      Tano K, Yasuda S, Kuroda T, Saito H, Umezawa A, Sato Y.
    • Journal Title

      PLoS One

      Volume: 9 Issue: 10 Pages: e110496-e110496

    • DOI

      10.1371/journal.pone.0110496

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] ヒトiPS細胞由来網膜色素上皮細胞中に混入する不死化細胞検出法の開発とその性能評価2016

    • Author(s)
      黒田拓也,安田智,松山さと子,髙田のぞみ,中島啓行,草川森士,梅澤明弘,松山晃文,川真田伸,佐藤陽治
    • Organizer
      第15回日本再生医療学会総会
    • Place of Presentation
      大阪国際会議場
    • Year and Date
      2016-03-17
    • Related Report
      2015 Annual Research Report
  • [Presentation] デジタル軟寒天コロニー形成試験を利用した再生医療製品の品質評価2016

    • Author(s)
      草川森士,安田智,黒田拓也,川真田伸,佐藤陽治
    • Organizer
      第15回日本再生医療学会総会
    • Place of Presentation
      大阪国際会議場
    • Year and Date
      2016-03-17
    • Related Report
      2015 Annual Research Report
  • [Presentation] Validation of in vitro methods for detection of immortalized cells in retinal pigment epithelial cells derived from human induced pluripotent stem cells2015

    • Author(s)
      Takuya Kuroda, Satoshi Yasuda, Satoko Matsuyama, Nozomi Takada, Hiroyuki Nakashima, Shinji Kusakawa, Akihiro Umezawa, Akifumi Matsuyama, Shin Kawamata, Yoji Sato
    • Organizer
      World stem cell summit15
    • Place of Presentation
      アトランタ
    • Year and Date
      2015-12-10
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Digital analysis of soft agar colony formation for highly sensitive detection of tumorigenic cellular impurities in human cell-processed therapeutic products2015

    • Author(s)
      Shinji Kusakawa, Satoshi Yasuda, Takuya Kuroda, Shin Kawamata, Yoji Sato
    • Organizer
      World stem cell summit15
    • Place of Presentation
      アトランタ
    • Year and Date
      2015-12-10
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] iPS細胞由来網膜色素上皮細胞の安全性評価ー造腫瘍性試験のデザイン策定についてー2015

    • Author(s)
      川真田伸,郷正博,金村星余,鹿村真之,西下直希,佐藤陽治,黒田拓也,草川森士,安田智,高橋政代,酒井徳子,森永千桂子,万代道子
    • Organizer
      第14回日本再生医療学会総会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2015-03-19 – 2015-03-21
    • Related Report
      2014 Research-status Report
  • [Presentation] ヒト多能性幹細胞由来再生医療製品に残存する未分化細胞をダイレクトに検出する方法の開発2015

    • Author(s)
      田埜慶子,安田智,黒田拓也,斎藤博久,梅澤明弘,佐藤陽治
    • Organizer
      第14回日本再生医療学会総会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2015-03-19 – 2015-03-21
    • Related Report
      2014 Research-status Report
  • [Presentation] 軟寒天コロニー形成試験を応用した再生医療製品に混在する悪性形質転換細胞の高感度検出法2015

    • Author(s)
      草川森士,安田智,黒田拓也,川真田伸,佐藤陽治
    • Organizer
      第14回日本再生医療学会総会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2015-03-19 – 2015-03-21
    • Related Report
      2014 Research-status Report
  • [Presentation] iPS細胞由来移植細胞の品質・安全性について2014

    • Author(s)
      黒田拓也
    • Organizer
      第58回日本薬学会関東支部大会
    • Place of Presentation
      昭和薬科大学
    • Year and Date
      2014-10-04
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] QX100 Droplet Digital PCRを用いたヒトiPS細胞由来分化細胞中における混入未分化iPS細胞の高感度検出法の開発2014

    • Author(s)
      黒田拓也
    • Organizer
      第1回 バイオ・ラッド デジタルPCRユーザーフォーラム
    • Place of Presentation
      天王洲アイルセントラルタワー
    • Year and Date
      2014-07-17
    • Related Report
      2014 Research-status Report
    • Invited
  • [Book] In vitro detection of residual undifferentiated cells in retinal pigment epithelial cells derived from human induced pluripotent stem cells.2014

    • Author(s)
      Takuya Kuroda, Satoshi Yasuda, Yoji Sato
    • Total Pages
      10
    • Publisher
      Springer
    • Related Report
      2014 Research-status Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

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