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Analysis of collected primitive hemopoietic stem cells and its clinical application

Research Project

Project/Area Number 04404086
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  Jichi Medical School, Professor, 医学部, 教授 (60048965)

Co-Investigator(Kenkyū-buntansha) GUNJI Yuji  Jichi Medical School.Assistant Professor, 医学部, 講師 (90245043)
HATAKE Kiyohiko  Jichi Medical School, Associate Professor, 医学部, 助教授 (80192699)
SHIMIZU Ritsuko  Jichi Medical School, Instructor, 医学部, 助手 (40226262)
IMAGAWA Shigehiko  Jichi Medical School, Assistant Professor, 医学部, 講師 (60231164)
KOMATSU Norio  Jichi Medical School, Assistant Professor, 医学部, 講師 (50186798)
角田 純一  自治医科大学, 医学部, 助手 (20245054)
室井 一男  自治医科大学, 医学部, 講師 (50190939)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥25,000,000 (Direct Cost: ¥25,000,000)
Fiscal Year 1994: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1993: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1992: ¥16,000,000 (Direct Cost: ¥16,000,000)
Keywordshemopoietic stem cell / hemopoietic factor / receptor / megakaryocytic cell line / tyrosine phosphorylation / erythropoietin receptor / hemopoietic stem cell disorders / apoptosis / チロシンフォスファターゼ
Research Abstract

1) Lineage- c-kit+ Sca-1+ cells collected from normal mouse bone marrow (BM) cells contain most primitive hemopoietic stem cells, containing one hemopoietic progenitor cell from 2 cells, whereas 100 cells could rescue lethally irradiated mice.
2) Human CD34+, c-kit + CD33 + cells were maintained their colony forming ability when cultured on a feeder layr derived from normal human BM cells beyond 6 weeks. Human umbilical cord blood cells taken from abortive delivery contained more primitive stem cells than those from full term delivery.
3) From a megakaryocytic cell line UT-7, we established an erythropoietin (EP) dependent subclone, which expressed abundant EP receptors EPR.EP tyrosine-phosphorylated various kinase proteins, among which phospholipase-Cgamma1 was phosphorylate, suggesting that inositol phospholipid pathway was also activated with EP.We also established a thrombopoietin (mpl-ligand, TP) dependent cell line, and differentiation of erythropoiesis and thrombopoiesis will be studied more in detail.
4) There are two types of EPR in normal erythroid cells, a full length (F) type and a truncated (T) type. F type prevents apotosis under the low concentration of EP,while T type does not. we suggested the role of c-jun oncogene along with the prevention of apoptosis.
5) We found that immature human stem cells did not express Fas, an apotosis related gene, while mature hemopoietic cells did. We are investigating the role of Fas in the myeloid hemopoiesis.
6) Hep3B cells, established from human hepatoma, produce EP under anoxic condition. In EP gene there is a CACCC element -60bp upstream from CAP-site, and GATA element -30kb upstream of CAP-site. Using antisense against either sequence, we found that the former serves as a positive regulatory element while the latter is a negative regulator element. We also found that in the nuclear extracts binding specifically to these elements.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Gunji,Y.,Nakamura,M.Miura,Y.,et al.: "Expression and function of adhesion molecules on human hematopoietic stem cells:CD34+LFA-1+ cells are more primitive than CD34+LFA-1+ cells." Blood. 80. 429-436 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okada,S.,Nakauchi,K.Miura,Y.,et al.: "In vivo and in vitro stem cell function of c-kit and Sca-1-positive murine hematopoietic cells." Blood. 80. 3044-3050 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okada,S.,Nagayoshi,K.Miura,Y.,et al.: "Sequential analysis of hematopoietic reconstitution achieved transplantation of hematopoietic stem cells." Blood. 81. 1720-1725 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Komatsu,N.,Yamamoto,M.Miura,Y.,et al.: "Establishment and characterization of an erythropoietin-dependent sabline,UT-7/Epo,derived from human leukemia cell line,UT-7." Blood. 82. 456-464 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Imagawa,S.,Izumi,T.Miura,Y.: "Positive and negative regulation of the erythropoietin gene." J.Biol.Chem.269. 9038-9044 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ren,HY.,Komatsu,N.Miura,Y.,et al.: "Erythropoietin induces tyrosine phosphorylation and activation of phospholipase C-γ 1 in a human erythropoietin-dependent cells line." J.Biol.Chem.269. 19633-19638 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hatake,K.,Tomizuka,H.Miura,Y.,et al.: "Mol.Biol.Haematopoiesis" Intercept Ltd., 7 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Gunji, Y., Nakamura, M., Hagiwara T., Hayakawa, K., Matsushita, H., Osawa, H., Nagayoshi, K., Nakauchi, H., Yanagisawa, M., Miura, Y., Suda, T.: "Expression and function of adhesion molecules of human hematopoietic stem cells : CD34^+LFA-1^- cells are more primitive than CD34^+LFA-1^+ cells." Blood. 80. 429-436 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Okada, S., Nagayoshi, K., Nakauchi, H., Nishikawa, S-I., Miura, Y., Suda, T.: "Sequential analysis of hematopoietic reconstitution achieved by transplantation of hematopoietic stem cells." Blood. 81. 1720-1725 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Komatsu, N., Yamamoto, M., Fujita, H., Miwa, A., Hatake, K., Endo, T., Okano, H., Katsube, T., Fukumaki, Y., Sassa, S., Miura, Y.: "Extablishment and characterization of an erythropoietin-dependent subline, UT-7/Epo, derived from human leukemia cell line, UT-7." Blood. 82. 456-464 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Gunji, Y., Nakamura, M., Osawa, H., Nagayoshi, K., Miura, Y., Yanagisawa, M., Suda, T.: "Human primitive hematopoietic progenitor cells are more enriched in KIT^<low> cells than in KIT^<high> cells." Blood. 82. 3283-3289 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Muroi, K., Suda T., Nakamura, M., Okada, S., Nojiri, H., Amemiya, Y., Miura, Y., Hakomori, S-I.: "Expression of sialosy-Tn in colony-forming unit-erythroid, erythroblasts, B cells, and a subset of CD4^+ cells." Blood. 83. 84-91 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Imagawa, S., Izumi, T., Miura, Y.: "Positive and negative regulation of the erythropoietin gene." J.Biol.Chem.269. 9038-9044 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ren, H-Y., Komatsu, N., Shimizu, R., Okada, K., Miura, Y.: "Erythropoietin induces tyrosine phosphorylation and activation of phospholipase C-E_i1 in a human erythropoietin-dependent cells line." J.Biol.Chem.269. 19633-19638 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Muroi,k.,Miura,Y.,et al.: "Expression of sialosyl‐Tn in colony‐forming unit‐erythroid,erythroblasts,B cells,and a subset of CD4+ cells." Blood. 83. 84-91 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ohtsuki,T.,Miura,Y.,et al.: "Alteration of the proteoglycan form of macrophage colony‐stimulating factor produced by a human stromal line‐‐‐‐." Experimental Hematology. 22. 366-369 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Imagawa,S.,Izumi,T.,Miura,Y.: "Positive and negative regulation of the erythropoietin gene." The Journal of Biological Chemistry. 269. 9038-9044 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ohtsuki,T.,Ikeda,M.,Miura,Y.,et al.: "Quantitative analysis of the two macrophage colony‐stimu‐lating factor mRNA expressed in a human stromal cell‐‐‐." Biochimica et Biopnysica Acta. 1222. 141-146 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ren,HU.,Komatsu,N.Miura,Y.,et al.: "Erythropoietin induces tyrosine phosphorylation and activation of phospholipase C‐γ1 in a human erythropoietin‐dependent cell line." The Journal of Biological Chemistry. 269. 19633-19638 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Suzuki,T.,Muroi,K.,Miura.Y.,et al.: "Characterization of enriched CD34‐positive cells from healthy volunteers and from patients treated with‐‐‐‐." Stem Cells. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Hatake,K.,Miura,Y et al.: "The presence of apoptosis in refractory anemia of myelodysplasia." Intercept Ltd, 7 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Miura,N.,Okada,S.,Miura,Y.,et al.: "Rat stem cell factor and IL-6 preferentially support the proliferation of c-kit-positive murine hemopoietic cells rather than their differentiation." Experimental Hematology. 21. 143-149 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Okada,S.,Nagayoshi,K.Miura,Y.,et al.: "Sequential analysis of hematopoietic reconstitution achieved by transplan-tation achieved by transplantation of hematopoietic stem cells." Blood. 81. 1720-1725 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Komatsu,N.,Yamamoto,M.Miura,Y.,et al.: "Establishment and characterization of an erythropoietin-dependent subline,UT-7/Epo,derived from human leukemia cell line,UT-7." Blood. 82. 456-464 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Moroi,M.,Ohnuma,M.,Komatsu.N.,Miura,Y.,: "Pborbol ester treatment of two megakaryoblastic cell lines,CMK and UT-7,induces specific protein composition changes with non-coincident time-courses." Biochimica et Biophysica Acta. 1117. 139-146 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Gunji,Y.,Nakamura,M.,Miura,Y.,et al.: "Human Primitive hematopoietic progenitor cells are more enriched in KIT low cells than in KIT high cells." Blood. 82. 3283-3289 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Muroi,K.,Suda,T.,Miura,Y.,et al.: "Expression of sialosyl-Tn in colony-forming unit-erythroid,erythroblasts B cells,and a subset of CD4+ cells." Blood. 83. 84-91 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Hatake,K.,Tomizuka,H.,Miura,Y.,et al.: "The presence of apoptosis in refractory anemia of myelodysplasia." Molecular Biology of Haematopoiesis. 7 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Gunji,Y.: "Expression and function of adhesion molecul on human hematopoietic stem cells:CD34^+LFA-1^+ cells." Blood. 80. 429-436 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Miura,N.: "Rat stem cell factor and lL-6 preferentially support the proliferation of c-kit-positive murine hemopoietic cells rather than their differentiation." Exp.Hematol.21. 143-149 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Okada,S.: "In vivo and in vitro stem cell function of c-kit and Sca-1-positlive murine hemopoietic cells." Blood. 80. 3044-3050 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Komatsu,N.: "Erythropoietin rapidly induces tyrosine phosphorylation in the human erythropoietin-dependent cell line,UT-7." Blood. 80. 53-59 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nakamura,Y.: "A truncated erythropoietin receptor that fails to prevent programmed cell death of erythroid cells." Sciince. 257. 1138-1141 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Shimomura,S.: "First continuous propagation of B19 parvovirus in a cell line." Blood. 79. 18-24 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Miura,T.,Ema,H.,Suda,T.,Nagayoshi,K.,Nakauchi,H.: "Myelodysplastic Syndrome & Cytokines" Elsevier Science Publishers B.V., 10 (1991)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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