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Isolation and characterization of somatic stem cells from human deciduous tooth-derived dental pulp cells by genetic engineering techniques

Research Project

Project/Area Number 18K09839
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 57070:Developmental dentistry-related
Research InstitutionKagoshima University

Principal Investigator

Inada Emi  鹿児島大学, 医歯学域鹿児島大学病院, 講師 (30448568)

Co-Investigator(Kenkyū-buntansha) 佐藤 正宏  鹿児島大学, 総合科学域総合研究学系, 教授 (30287099)
齊藤 一誠  新潟大学, 医歯学系, 准教授 (90404540)
野口 洋文  琉球大学, 医学(系)研究科(研究院), 教授 (50378733)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords遺伝子工学的手法 / 乳歯歯髄幹細胞 / 体性幹細胞
Outline of Final Research Achievements

Based on our previous hypothesis [human deciduous tooth-derived dental pulp cells (HDDPCs) with high alkaline phosphatase (ALP) activity and enriched with OCT-3/4 and SOX2 tend to be easily reprogrammed into induced pluripotent stem cells when they are transfected with vectors carrying Yamanaka’s reprogramming factors], we isolated two or more (~10) ALP-positive (HDDPC-ALP(+)) and negative (HDDPC-ALP(-)) cells in HDDPCs for molecular profiling. Consequently, we demonstrated that HDDPC-ALP(+) cells expressed both ALP and OCT-3/4. In contrast, HDDPC-ALP(-) cells expressed SOX2, but not ALP and OCT-3/4. These findings indicate that there are various types of somatic stem cells with different pluripotent cell-specific gene expression profiles in HDDPCs.

Academic Significance and Societal Importance of the Research Achievements

幹細胞を特徴付けるマーカーの多くはOCT-3/4、SOX2等の核内転写因子やALP等の細胞内因子であり、これらを発現する幹細胞は生きた形では細胞表面分子を標的としたFACS等による細胞捕捉で単離・濃縮できない。本研究では、HDDPCから幹細胞特異的遺伝子を発現する少数個の細胞を単離し、微量なmRNAを調製した。これを基にする分子生物学的解析から、個々の細胞における遺伝子発現様式を解明することができた。
本法により未知の分子マーカー発見の可能性があること、少数個の細胞で特定の遺伝子発現プロファイル作成が可能となることが期待され、HDDPCに含まれる幹細胞の特性をあぶりだす点で学術的意義は高い。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (14 results)

All 2021 2020 2019 2018

All Journal Article (12 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 12 results,  Open Access: 12 results) Presentation (2 results)

  • [Journal Article] RNA analysis based on a small number of manually isolated fixed cells (RNA-snMIFxC) to profile stem cells from human deciduous tooth-derived dental pulp cells2021

    • Author(s)
      Inada E, Saitoh I, Kubota N, Iwase Y, Kiyokawa Y, Noguchi H, Yamasaki Y, Sato M
    • Journal Title

      Biological Procedure Online

      Volume: -

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] piggyBac-Based Non-Viral In Vivo Gene Delivery Useful for Production of Genetically Modified Animals and Organs.2020

    • Author(s)
      Sato M, Inada E, Saitoh I, Watanabe S, Nakamura S.
    • Journal Title

      Pharmaceutics

      Volume: 12 Issue: 3 Pages: 277-277

    • DOI

      10.3390/pharmaceutics12030277

    • Related Report
      2020 Annual Research Report 2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Drug-induced naive iPS cells exhibit better performance than primed iPS cells with respect to the ability to differentiate into pancreatic β-cell lineage2020

    • Author(s)
      Kiyokawa Y, Sato M, Noguchi H, Inada E, Iwase Y, Kubota N, Sawami T, Terunuma M, Maeda T, Hayasaki H, Saitoh I
    • Journal Title

      Journal of Clinical Medicine

      Volume: 9 Issue: 9 Pages: 2838-2838

    • DOI

      10.3390/jcm9092838

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of a novel, pipette tip-aided cell cloning method for effective isolation of genome-edited porcine cell2020

    • Author(s)
      Sato M, Saitoh I, Akasaka E, Inada E
    • Journal Title

      OBM Genetics

      Volume: 5 Issue: 1 Pages: 16-16

    • DOI

      10.21926/obm.genet.2101126

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Increased Expression of Cell Surface SSEA-1 is Closely Associated with Na?ve-Like Conversion from Human Deciduous Teeth Dental Pulp Cells-Derived iPS Cells2019

    • Author(s)
      Inada Emi、Saitoh Issei、Kubota Naoko、Iwase Yoko、Murakami Tomoya、Sawami Tadashi、Yamasaki Youichi、Sato Masahiro
    • Journal Title

      International Journal of Molecular Sciences

      Volume: 20 Issue: 7 Pages: 1651-1651

    • DOI

      10.3390/ijms20071651

    • Related Report
      2019 Research-status Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] In Vivo Piggybac-Based Gene Delivery towards Murine Pancreatic Parenchyma Confers Sustained Expression of Gene of Interest.2019

    • Author(s)
      Sato M, Inada E, Saitoh I, Nakamura S, Watanabe S
    • Journal Title

      International Journal of Molecular Science

      Volume: 20 Issue: 13 Pages: 3116-3116

    • DOI

      10.3390/ijms20133116

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Potential for Isolation of Immortalized Hepatocyte Cell Lines by Liver-Directed In Vivo Gene Delivery of Transposons in Mice.2019

    • Author(s)
      Sato M, Saitoh I, Inada E, Nakamura S, Watanabe S
    • Journal Title

      Stem Cells International

      Volume: 2 Pages: 5129526-5129526

    • DOI

      10.1155/2019/5129526

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] piggyBac Transposon-Based Immortalization of Human Deciduous Tooth Dental Pulp Cells with Multipotency and Non-Tumorigenic Potential.2019

    • Author(s)
      Inada E, Saitoh I, Kubota N, Iwase Y, Kiyokawa Y, Shibasaki S, Noguchi H, Yamasaki Y, Sato M
    • Journal Title

      International Journal of Molecular Science

      Volume: 20 Issue: 19 Pages: 4904-4904

    • DOI

      10.3390/ijms20194904

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Repeated human deciduous tooth-derived dental pulp cell reprogramming factor transfection yields multipotent intermediate cells with enhanced iPS cell formation capability2019

    • Author(s)
      Soda Miki、Saitoh Issei、Murakami Tomoya、Inada Emi、Iwase Yoko、Noguchi Hirofumi、Shibasaki Shinji、Kurosawa Mie、Sawami Tadashi、Terunuma Miho、Kubota Naoko、Terao Yutaka、Ohshima Hayato、Hayasaki Haruaki、Sato Masahiro
    • Journal Title

      Scientific Reports

      Volume: 9 Issue: 1 Pages: 1490-1490

    • DOI

      10.1038/s41598-018-37291-2

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Timing of CRISPR/Cas9-related mRNA microinjection after activation as an important factor affecting genome editing efficiency in porcine oocytes2018

    • Author(s)
      Sato M, Maeda K, Koriyama M, Inada E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai, T, Watanabe S, Miyoshi K
    • Journal Title

      Theriogenology

      Volume: 108 Pages: 29-38

    • DOI

      10.1016/j.theriogenology.2017.11.030

    • NAID

      120007099790

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Intrapancreatic Parenchymal Cell Transplantation as a Possible Model for the Development of a Cell-based Therapy for Type I Diabetes Mellitus2018

    • Author(s)
      Sato M, Inada E, Nakamura S, Saitoh I
    • Journal Title

      OBM Transplantation

      Volume: 2 Issue: 3 Pages: 1-1

    • DOI

      10.21926/obm.transplant.1803016

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Advance in drug-free acquisition of gene-engineered mammalian cells: A mini-review.2018

    • Author(s)
      Sato M, Inada E, Saitoh I, Yamasaki Y, Watanabe S
    • Journal Title

      Current Topics in Biotechnology

      Volume: 9 Pages: 1-9

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 免疫染色された少数細胞の簡便な取得に基づくヒト歯髄細胞の遺伝子発現解析2020

    • Author(s)
      稲田絵美, 齊藤一誠, 清川裕貴, 早崎治明, 山崎要一
    • Organizer
      第58回日本小児歯科学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 初期胚特異的糖鎖抗原SSEA-1は乳歯歯髄細胞由来iPS細胞の高度未分化状態を特定するマーカーとして有用である2018

    • Author(s)
      稲田絵美, 齊藤一誠, 窪田直子, 村上智哉, 澤味 規, 松枝一成, 早崎治明, 山崎要一
    • Organizer
      第56回日本小児歯科学会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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