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1991 Fiscal Year Final Research Report Summary

Isolation concentration, and preservation of hemopoietic stem cells and its clinical

Research Project

Project/Area Number 01440049
Research Category

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

Allocation TypeSingle-year Grants
Research Field Hematology
Research InstitutionJichi Medical School

Principal Investigator

MIURA Yasusada  Department of Medicine, JIchi Medical School, Professor, 医学部, 教授 (60048965)

Co-Investigator(Kenkyū-buntansha) IMAGAWA Shigehiko  Jichi Medi. School, Dept. Medi., Assistant, 医学部, 助手 (60231164)
MIWA Akiyoshi  Jichi Medi. School, Dept. Medi., Assistant Prof., 医学部, 講師 (40200224)
MUROI Kazuo  Jichi Medi. School, Dept. Medi., Assistant, 医学部, 助手 (50190939)
KOMATSU Norio  Jichi Medi. School, Dept, Medi., Assistant, 医学部, 助手 (50186798)
SUDA Toshio  Jichi Medi. School, Dept. Medi., Associate Prof., 医学部, 助教授 (60118453)
Project Period (FY) 1989 – 1991
Keywordshemopoietic stem cells / hemopoietic growth factors / monoclonal antibody / CD34 / c-kit / mucopolysaccharide / spleen colony / anti-leukemic cell antibody
Research Abstract

The purpose of this research is to sort out and concentrate hemopoietic stem cells, which is only a minor population of hemopoietic tissue, to study their reactivity to growth factors, and finally to investigate the clinical applications.
The main results are as follows :
1) We sorted CD34^+CD33^- cell fraction from normal human bone marrow cells by a fluorescence activated cell sortor (FACS), which contain the most primitive stem cells. After treating these cells with IL-3, they turned into CD34^- CD33^+ cells and then were induced to mature blood cells by the addition of lineage specific cytokines such as IL-5 and G-CSF.
2) Murine hemopoietic cells were sorted by various monoclonal antibodies using a FACS. The c-kit^+, SCA-1^+ lineage^- cells were found to contain highly enriched primitive stem cells. They could reconstitute murine hemopoiesis when injected into lethally irradiated mice.
3) Sorted stem cells were of middle-size with round, immature nuclei and basophilic cytoplasm occasionally with some granules.
4) We established a riiegakaryocytic cell line, UT-7, which is completely dependent on various cytokines such as IL-3, GM-CSF, erythropoietin or IL-6. We investigated the appearance of erythropoietin receptors and followed the signal transduction pathways. A mucopolysaccharide was extracted and purified from this cell line cells, which induced the "differentiation" of original UT-7 cells when added in vitro.
5) Several anti-sugar chain monoclonal antibody were became available. Some of them were shown to recognize human leukemic cells, but not the normal sorted stem cells. This phenomenon could be applied on the purging of leukemic cells from patient bone marrow at the time of autologous bone marrow transplantation.
As a conclusion, our research projects yielded many valuable results published in leading international journals, and could be useful for both basic and clinical study on the hemopoiesis.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Suda,T.Okada,S.,Suda,J.,Miura,Y.,et al.: "A stimulatory effect of recombinant murine interleukin-7(IL-7)on-B-cell colony formation and inhibitory effect of IL-1 α." Blood. 74. 1936-1941 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ema,H.,Suda,T.,Miura,Y.,Nakauchi,H.: "Colony formation of clone-sorted human hematopoietic progenitors." Blood. 75. 1941-1946 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Gunji,Y.: "Support of early B-cell differentiation in mouse fetal liver by stromal cells and interleukin-7." Blood. 77. 2612-2617 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Komatsu.,N.: "Establishment and characterization of a human leukemic cell line with megakaryocytic features:Dependency on granulocyte-macrophage for colony-stimulating factor,interleukin 3,or erythropoietin growth and survival." Cancer Research. 51. 341-348 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Okada,S.,Nakauchi,H.,Nagayoshi,K.,Nishikawa,S.,Nishikawa,S.,Miura,Y.Suda,T.: "Enrichment and characterization of murine hematopoietic stem cells that express c-kit molecule." Blood. 78. 1706-1712 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Muroi,K.,Suda,T.,Nojiri,T.,Ema,H.,Amemiya,Y.,Miura,Y.,Hakomori,S.,et al.: "Reactivityprofiles of leukemic myeloblasts with monoclonal antibodies directed to sialosyl-Le^x and other lacto-series type 2 chain antigens:Absence of reactivity with normal hematopoietic progenitor cells." Blood. 79. 713-719 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miura,Y.,Komatsu,N.,Suda,T.: "(Molecular biology and differentiation of megakaryocytes)Growth and differentiation of two human megakaryoblastic cell lines;CMK and UT-7." Wiley-Liss,New York, 12 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Miura,Y.: "(Myelodysplastic Syndrome and Cytokines)Differentiation pathways of clone-sorted CD34-positive cells." Elsevier Science Publishers, 39-48 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suda, T. Okada, S., Suda, J., Miura, Y., et al.: "A stimulatory effect of recombinant murine interleukin-7 (IL-7) on B-cell colony formation and inhibitory effect of IL-1 alpha." Blood. 74. 1936-1941 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ema, H., Suda, T., Miura, Y., Nakauchi, H.: "Colony formation of clone-sorted human hematopoietic progenitors." Blood. 75. 1941-1946 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Gunji, Y.: "Support of early B-cell differentiation in mouse fetal liver by stromal cells and interleukin-7." Blood. 77. 2612-2617 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Komatsu, . N.: "Establishment and characterization of a human leukemic cell line with megakaryocytic features : Dependency on granulocyte-macrophage colony-stimulating factor, interleukin 3, or erythropoietin for growth and survival." Cancer Research. 51. 341-348 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Okada, S., Nakauchi, H.: "Enrichment and characterization of murine hematopoietic stem cells that express c-kit molecule." Blood. 78. 1706-1712 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Muroi, K., Suda, T.: "Reactivity profiles of leukemic myeloblasts with monoclonal antibodies directed to sialosyl-Le^x and other lacto-series type 2 chain antigens : Absence of reactivity with normal hematopoietic progenitor cells." Blood. 79. 713-719 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miura, Y., Komatsu, N., Suda, T.: "Growth and differentiation of two human megakaryoblastic cell lines ; CMK and UT-7." Molecular biology and differentiation of megakaryocytes. Wiley-Liss. (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Miura, Y.: "Differentiation pathways of clone-sorted CD34-positive cells." Myelodysplastic Syndrome and Cytokines. Elsevier Science Publishers. (1991)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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