• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Studies on regulatory mechanisms of megakaryocytepoiesis using the cell line grown in protein-free culture.

Research Project

Project/Area Number 05670686
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Pediatrics
Research InstitutionYokohama City University School of Medicine

Principal Investigator

SASAKI Hideki  Yokohama City University, School of Medicine, Associate Professor, 医学部, 助教授 (50106316)

Co-Investigator(Kenkyū-buntansha) MATSUYAMA Shusuke  Yokohama City University, School of Medicine, Professor, 医学部, 教授 (20045983)
FUNABIKI Tetsunori  Yokohama City University, School of Medicine, Instructor, 医学部, 助手 (20264616)
IKUTA Koichiro  Yokohama City University, School of Medicine, Assistant Professor, 医学部, 講師 (80159590)
甲斐 純夫  横浜市立大学, 医学部, 助手 (10233644)
井上 達  横浜市立大学, 医学部, 助教授 (50100110)
Project Period (FY) 1993 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
Keywordsmegakaryocyte / cell line / stem cell / cytokine / thrombopoietin / TGF / 細胞株 / 無蛋白培養 / サイトカイン
Research Abstract

We examined the effects of cytokines on the growth of murine megakaryocytic cell lines, L8057 and L8057Y5, which had been maintained in protein-free culture. In serum-free methylcellulose culture, the addition of rhEPO,rmIL-3, rhIL-6, rmLIF,or rmGM-CSF enhanced colony growth of L8057Y5 cells mostly in dose-dependent fashion Also, colony growth of L8057 cells was enhanced by these cytokines, but to a lesser extent. The expression of mRNAs for EPO-R,IL-6R,gp130, IL-3R subunits were constitutively expressed in both lines. We also tested the effect of rmTpo, which was newly cloned in 1994, on the megakaryocyte progenitors in marrow. The megakaryocyte colony growth was observed by the stimulation of sole rmTpo. The addition of rmIL-3 with rmTpo showed increase not only in the total number, but in the frequencies of large colonies and of mixed-type colonies. Also, rhEpo and GM-CSF showed synergistic effects with rmTpo on the colony formation. Colony formation by BFU-Es and CFU-GMs was not significantly reduced by the addition of rhTGF-beta_1 ; however, the growth of CFU-Mks, stimulated by rmIL-3, rmTPO,or both cytokines, was markedly inhibited, which suggest that TGF-beta is a potent negative-regulator in megakaryocytepoiesis.
rmTpo also enhanced the growth of L8057 cells mostly in a concentration-dependent fashon as observed in the growth of CFU-Mk. RT-PCR revealed the costitutive expression of Mpl-mRNA in L8057 cells. These results suggest that L8057 and L8057Y5 cell lines have the characteristics of megakaryoblastic cells in their biological responses to cytokines, and that the growth-enhancing effects of these cytokines on the cell lines may be achieved through specific receptors. For totally clarifying the regulation mechanisms of thrombopoiesis, it should be promissing way to investigate signaling cascades for every cytokines including Tpo in megakaryoblastic cells. Again our findings validate the usefulness of L8057 cells for these purposes.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Y. Ishida et al.: "Biological and biochemical characteristics of murine megakaryoblastic cell line L8057." Experimental Hematology. 21. 289-298 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] S. Taniguchi et al.: "Hemopoietic stem-cell compartment of the SCID mouse: Double-exponential survive curve after γ irradiation." Proceeding of National Academy of Science USA. 90. 4354-4358 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] H. Sasaki et al.: "The clinical course of acquired aplastic anemia in childhood; a retrospective study" Internatinal Journal of Hematology. 60. 239-249 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 佐々木秀樹ほか: "小児再生不良性貧血における造血前駆細胞動態とstem cell factorの増殖促進 作用についての解析" 日本小児科学会雑誌. 98. 903-909 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] H.Sasaki et al.: "Effects of erythropoietin, IL-3,IL-6,and LIF on a murine megakaryoblastic cell line: Growth enhancement and expression of receptor mRNAs." Leukemia Research. 19. 95-102 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Inoue et al.: "Survival of spleen colony-forming units(CFU-S)of irradiated bone-marrow cells in mice-Evidence for the existance of a radio-resistant subfraction." Experimental Hematology. 23. 1296-1300 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Y.Ishida, J.Levin, G.Baker, PE.Stenberg, Y.Yamada, H.Sasaki, T.Inoue: "Biological and biochemical characteristics of murine megakaryoblastic cell line L8057." Experimental Hematol.21. 289-298 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] S.Taniguchi, Y.Hirabayashi, T.Inoue, M.Kanisawa, H.Sasaki, K.Komatsu, KJ.Mori: "Hemopoietic stem-cell compartment of the SCID mouse : Double-exponential survive curve after gamma irradiation." Proc.Natl.Acad.Sci.USA. 90. 4345-4358 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] H.Sasaki, K.Ikuta, T.Okuyama, T.Funabiki, Y.Kajigaya, S.Kai, H.Sekiguchi, N.Hanzawa, H.Sumita, H.Takahashi, K.Fujioka, M.Matsuda, S.Matsuyama: "The clinical course of acquired aplastic anemia in childhood ; a retrospective study." Int.J.Hematol.60. 239-249 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] H.Sasaki, Y.Hirabayashi, T.Ishibashi, T.Inoue, M.Matsuda, S.Kai, K.Ikuta, K.Yokoyama, T.Yokota, Y.Maruyama, S.Matsuyama: "Effects of erythropoietin, IL-3, IL-6, and LIF on a murine megakaryoblastic cell line : Growth enhancement and expression of receptor mRNAs." Leukemia Research. 19. 95-102 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] T.Inoue, Y.Hirabayashi, H.Mitsui, H.Sasaki, E.P.Cronkite, J.E.Bullis, Jr, V.P.Bond, K.Yoshida: "Survival of spleen colony-forming units (CFU-S) of irradiated bone-marrow cells in mice : Evidence for the existance of a radio-resistant subfraction." Exp.Hematol.23. 1296-1300 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Y.Ishida et al: "Biological and biochemical characteristics of murine megakaryoblastic cell line L8057" Experimental Hematol. 21. 289-298 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] S.Taniguchi et al.: "Hemopoietic stem-cell compartment of the SCID mouse:Double-exponential survive curve after γ irradiation." Proc.Natl.Acad.Sci.USA. 90. 4354-4358 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Sasaki et al.: "The clinical course of acquired aplastic anemia in childhood;a retrospective study." Int.J.Hematol.60. 239-249 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Sasaki et al.: "Effects of erythropoietin,IL-3,IL-6,and LIF on a murine megakaryoblastic cell line:Growth enhancement and expression of receptor mRNAs." Leukemia Research. 19. 95-102 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Inoue et al.: "Survival of spleen colony-forming units(CFU-S)of irradiated bone-marrow cells in mice:Evidence for the existace of a radio-resistant subfraction." Exp.Hematol.23. 1296-1300 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 佐々木秀樹 ほか: "小児再生不良性貧血における造血前駆細胞動態とstem cell factorの増殖促進作用についての解析" 日本小児科学会雑誌. 98. 903-909 (1994)

    • Related Report
      1995 Annual Research Report

URL: 

Published: 1993-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi