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Toward the establishment of culture system to restore in vivo phenotypes of podocytes

Research Project

Project/Area Number 24591190
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Kidney internal medicine
Research InstitutionNiigata University

Principal Investigator

YAOITA Eishin  新潟大学, 医歯学系, 准教授 (00157950)

Co-Investigator(Kenkyū-buntansha) YOSHIDA Yutaka  新潟大学, 医歯学系, 講師 (40182795)
TAKEUCHI Kousei  愛知医科大学, 医学部, 教授 (90206946)
YAMAMOTO Tadashi  新潟大学, 医歯学系, 教授 (30092737)
Project Period (FY) 2012-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsポドサイト / 培養 / 継代 / トリプシン / ヘパリン / ビタミンA / 細胞外基質 / 細胞密度 / 単離糸球体 / ネフリン / ポドシン / コラゲン / 胎仔牛血清 / 足細胞 / 硫酸化 / デキサメサゾン / トランスクリプトーム / 腎臓 / 遺伝子発現
Outline of Final Research Achievements

Glomerular epithelial cells in the kidney (podocytes) play critical roles in maintenance of glomerular structure and glomerular ultrafiltration. Establishment of culture system remaining in vivo phenotypes of podocytes is essential for studies of podocytes, although podocytes drastically decrease podocyte-specific gene expressions with time in the primary culture. Of these, decrease in nephrin was the most evident. In this study, we evaluated effect of various culture conditions on the gene expression in primary culture of podocytes to improve phenotypes of cultured cells to restore in vivo phenotypes. Consequently, subculture on day 3 of culture of isolated glomeruli, detachment with non-enzymatic solution, addition of heparin, decrease in fetal bovine serum, adequate cell density, and extracellular matrices enable expression of podocyte-specific genes at the level of in vivo. In addition, dexamethasone and all trans-retinoic acid induce cellular processes of cultured podocytes.

Report

(4 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • 2012 Research-status Report
  • Research Products

    (14 results)

All 2015 2014 2013 2012

All Journal Article (7 results) (of which Peer Reviewed: 5 results,  Open Access: 3 results,  Acknowledgement Compliant: 2 results) Presentation (7 results)

  • [Journal Article] Label-free quantitative proteomic analysis reveals strong involvement of complement alternative and terminal pathways in human glomerular sclerotic lesions.2015

    • Author(s)
      Zhang Y, Xu B, Kinoshita N, Yoshida Y, Tasaki M, Fujinaka H, Magdeldin S, Yaoita E, Yamamoto T.
    • Journal Title

      J Proteomics.

      Volume: 123 Pages: 89-100

    • DOI

      10.1016/j.jprot.2015.03.024

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Expression of tight junction protein claudin-1 in human crescentic glomerulonephritis.2014

    • Author(s)
      Koda R, Yoshino A, Imanishi Y, Kawamoto S, Ueda Y, Yaoita E, Kazama JJ, Narita I, Takeda T.
    • Journal Title

      Int J Nephrol

      Volume: 2014 Pages: 598670-598670

    • DOI

      10.1155/2014/598670

    • Related Report
      2014 Annual Research Report
    • Open Access
  • [Journal Article] Deep proteome mapping of mouse kidney based on OFFGel prefractionation reveals remarkable protein post- translational modifications.2014

    • Author(s)
      Magdeldin S, Yamamoto K, Yoshida Y, Xu B, Zhang Y, Fujinaka H, Yaoita E, Yates JR 3rd, Yamamoto T
    • Journal Title

      J Proteome Res.

      Volume: 13 Issue: 3 Pages: 1636-46

    • DOI

      10.1021/pr401122m

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] 糸球体足細胞の培養系2014

    • Author(s)
      矢尾板永信
    • Journal Title

      腎と透析

      Volume: 77 Pages: 406-411

    • Related Report
      2014 Annual Research Report
  • [Journal Article] Heparin increasing podocyte-specific gene expressions.2014

    • Author(s)
      Yaoita E, Yoshida Y, Nameta M, Zhang Y, Fujinaka H, Magdeldin S, Xu B, Yamamoto T.
    • Journal Title

      Nephrology (Carlton)

      Volume: 19 Issue: 4 Pages: 195-201

    • DOI

      10.1111/nep.12207

    • Related Report
      2013 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Profiling and annotation of human kidney glomerulus proteome2013

    • Author(s)
      Cui Z, Yoshida Y, Xu B, Zhang Y, Nameta M, Magdeldin S, Makiguchi T, Ikoma T, Fujinaka H, Yaoita E, Yamamoto T
    • Journal Title

      Proteome Sci

      Volume: 11 Issue: 1 Pages: 13-13

    • DOI

      10.1186/1477-5956-11-13

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Proteomic approach to human kidney glomerulus prepared by laser microdissection from frozen biopsy specimens: exploration of proteome after removal of blood-derived proteins2012

    • Author(s)
      Yoshida Y, Nameta M, Kuwano M, Zhang Y, Bo X, Magdeldin S, Cui Z, Fujinaka H, Yaoita E, Tomonaga T, Yamamoto T
    • Journal Title

      Proteomics Clin Appl

      Volume: 6 Issue: 7-8 Pages: 412-417

    • DOI

      10.1002/prca.201200016

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Presentation] ポドサイトにおける接着結合2014

    • Author(s)
      矢尾板 永信、西村 宏子、行田 正晃、山本 格
    • Organizer
      第57回日本腎臓学会学術総会
    • Place of Presentation
      横浜
    • Year and Date
      2014-06-04 – 2014-06-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] Profiling and annotation of human kidney glomerulus proteome2014

    • Author(s)
      吉田 豊、崔 珍玉、許 波、藤中 秀彦、矢尾板 永信、山本 格
    • Organizer
      第57回日本腎臓学会学術総会
    • Place of Presentation
      横浜
    • Year and Date
      2014-06-04 – 2014-06-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] 質量分析計と抗体の両者で同定された高信頼ヒト糸球体プロテオーム2014

    • Author(s)
      行田 正晃、吉田 豊、矢尾板 永信、許 波、張エイ、山本 格
    • Organizer
      第57回日本腎臓学会学術総会
    • Place of Presentation
      横浜
    • Year and Date
      2014-06-04 – 2014-06-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] 質量分析と免疫組織化学によるヒト糸球体プロテオームデータベースの比較2013

    • Author(s)
      行田正晃、吉田豊、矢尾板永信、許波、張エイ、山本格
    • Organizer
      第56回日本腎臓学会学術総会
    • Place of Presentation
      東京国際フォーラム(東京)
    • Related Report
      2013 Research-status Report
  • [Presentation] ラット足細胞初代培養におけるヘパリンの効果(2)2013

    • Author(s)
      矢尾板永信、吉田豊、山本格
    • Organizer
      第56回日本腎臓学会学術総会
    • Place of Presentation
      東京国際フォーラム(東京)
    • Related Report
      2013 Research-status Report
  • [Presentation] 糸球体単離時の尿細管混入が糸球体発現遺伝子発現の定量に与える影響について2013

    • Author(s)
      水谷栄介、矢尾板永信、山本格
    • Organizer
      第56回日本腎臓学会学術総会
    • Place of Presentation
      東京国際フォーラム(東京)
    • Related Report
      2013 Research-status Report
  • [Presentation] ラット足細胞初代培養におけるヘパリンの効果2012

    • Author(s)
      矢尾板 永信、山本 格
    • Organizer
      日本腎臓学会学術総会
    • Place of Presentation
      横浜
    • Related Report
      2012 Research-status Report

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Published: 2013-05-31   Modified: 2019-07-29  

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