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Analysis of side population cells as keratinocyte stem cells.

Research Project

Project/Area Number 16591102
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Dermatology
Research InstitutionEhime University

Principal Investigator

KAMEDA Kenji  Ehime University, Integrated Center for Sciences, lecturer, 総合科学研究支援センター, 講師 (60363264)

Co-Investigator(Kenkyū-buntansha) SHIRAKATA Yuji  Ehime University, Graduate School of Medicine, lecturer, 大学院医学系研究科, 講師 (50226320)
Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
Keywordsstem cells / SP-cells / cell sorter / keratinocyte
Research Abstract

Cultured skin is widely used for skin defects such as chronic ulcer, burns and so on. To develop gene therapy using cultured skin, it is necessary to introduce foreign genes into keratinocyte stem cells. Keratinocyte stem cells have been isolated using cell surface markers, however this method requires multiple steps for staining, and cell damage might occur because of the use of antibodies. Side population (SP) cells was firstly reported as stem cell isolation method for hematopoietic cells. The purpose of this study is to investigate whether SP cells exist in keratinocytes and SP cells are keratinocyte stem cells or not. Normal human skin was obtained from plastic surgery. Epidermis was separated from dermis by dispase treatment, single cells of keratinocytes were obtained by trypsin treatement. Keratinocytes were collected and cultured under serum free condition. Cells were stained with Hoechst 33342, then Hoechst Red and Blue expression was analyzed by cell sorter. SP cells exist in normal keratinocytes, and the population rate is 1-5% in total cells. Next we performed microarray analysis. SP cells as well as main population (MP) cells were sorted and total RNA was extracted. Using gene chip carrying 27000 genes, about 10 genes were significantly up-or down-regulated. Finally we investigated proliferative potency of SP cells. SP cells and MP cells were sorted and cultured separately. When cells reached subconfluent, cells were passaged. SP cells had higher proliferative potency by 3-4 passages, however MP cells grew well after 5 passages. These data indicate that SP cells might be the transit amplifying cells rather than stem cells, however further studies are needed.

Report

(4 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (15 results)

All 2007 2006

All Journal Article (15 results)

  • [Journal Article] Pre-B-cell leukemia transcription factor 1 is a major target of promyelocytic leukemia zinc-finger-mediated melanoma cell growth suppression.2007

    • Author(s)
      Shiraishi K, Yamasaki K, Nanba D, Inoue H, Hanakawa Y, Shirakata Y, Hashimoto K, Higashiyama S
    • Journal Title

      Oncogene 26

      Pages: 339-348

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Epiregulin, a member of the EGF family, is over-expressed in psoriati epidermis.2007

    • Author(s)
      Shirakata Y, Kishimoto J, Tokumar S, Yamasaki K, Hanakawa Y, Tohyama M, Sayama K, Hashimoto K
    • Journal Title

      J Dermatol Sci. 45

      Pages: 69-72

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects2007

    • Author(s)
      Yoshida T, Hakuba N, Morizane I, Fujita K, Cao F, Zhu P, Uchida N, Kameda K, Sakanaka M, Gyo K, HataR
    • Journal Title

      Neuroscience 145

      Pages: 923-930

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Pre-B-cell leukemia transcription factor 1 is a major target of promyelocytic leukemia zinc-finger-mediated melanoma cell growth suppression.2007

    • Author(s)
      Shiraishi K, Yamasaki K, Nanba D, Inoue H, Hanakawa Y Shirakata Y, Hashimoto K, Higashiyama S.
    • Journal Title

      Oncogene. 26

      Pages: 339-48

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Epiregulin, a member of the EGF family, is over-expresses in psoriatic epidermis.2007

    • Author(s)
      Shirakata Y, Kishimoto J, Tokumaru S, Yamasaki K, Hanakawa Y, Tohyama M, Sayama K, Hashimoto K.
    • Journal Title

      J Dermatol Sci. 45

      Pages: 69-72

    • NAID

      10019994988

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects.2007

    • Author(s)
      Yoshida T, Hakuba N, Morizane I Fujita K, Cao F, Zhu P, Uchida N, Kameda K, Sakanaka M, Gyo K, Hata R..
    • Journal Title

      Neuroscience. 145

      Pages: 923-930

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Hematopoietic stem cells prevent hair cell death after transient cochlear ischemia through paracrine effects2007

    • Author(s)
      Yoshida T, Hakuba N, Morizane I, Fujita K, Cao F, Zhu P, Uchida N, Kameda K, Sakanaka M, Gyo K, HataR.
    • Journal Title

      Neuroscience 145

      Pages: 923-930

    • Related Report
      2006 Annual Research Report
  • [Journal Article] New skin-equivalent model from de-epithelialized amnion membrane.2006

    • Author(s)
      Yang L, Shirakata Y, Shudou M, DailX, Tokumaru S, Hirakawa S, Sayama K, Hamuro J, Hashimoto K
    • Journal Title

      Cell Tissue Res. 326

      Pages: 69-77

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] TAK1 is essential for differentiation and the prevention of apoptosi in epidermis2006

    • Author(s)
      Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang. L, Sato S, Shizuo A, Hashimoto K
    • Journal Title

      J Biol Chem 281

      Pages: 22013-22020

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] NG2 proteoglycan-expressing microglia as multipotent neural progenitors in normal and pathologic brains.2006

    • Author(s)
      Yokoyama A, Sakamoto A, Kameda K, Imai Y, Tanaka J
    • Journal Title

      GLIA 53

      Pages: 754-768

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] New skin- equivalent model from de-epithelialized amnion membrane2006

    • Author(s)
      Yang L, Shirakata Y, Shudou M, DailX, Tokumaru S, Hirakv S, Sayama K, Hamuro J, Hashimoto K.
    • Journal Title

      Cell Tissue Res. 326

      Pages: 69-77

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] TAK1 is essential for differentiation and the prevention of apoptosis in epidermis.2006

    • Author(s)
      Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang L, Sato S, Shizu A, Hashimoto K.
    • Journal Title

      J Biol Chem. 281

      Pages: 22013-20

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] NG2 proteoglycan-expressing microglia as multipotent neural progenitors in normal and pathologic brains.2006

    • Author(s)
      Yokoyama A, Sakamoto A, Kameda K, Imai Y, Tanaka J.
    • Journal Title

      GLIA. 53

      Pages: 754-768

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] TAK1 is essential for differentiation and the prevention of apoptosis in epidermis.2006

    • Author(s)
      Sayama K, Hanakawa Y, Nagai H, Shirakata Y, Dai X, Hirakawa S, Tokumaru S, Tohyama M, Yang L, Sato S, Shizuo A, Hashimoto K
    • Journal Title

      J Biol Chem 281

      Pages: 22013-20

    • Related Report
      2006 Annual Research Report
  • [Journal Article] NG2 proteoglycan-expressing microglia as multipotent neural progenitors in normal and pathologic brains.2006

    • Author(s)
      Yokoyama A, Sakamoto A, Kameda K, Imai Y, Tanaka J.
    • Journal Title

      GLIA 53

      Pages: 754-768

    • Related Report
      2006 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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