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Development of the system to produce large scale generation of human platelets from cord blood CD34^+ cells in vitro

Research Project

Project/Area Number 18591076
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionSapporo Medical University

Principal Investigator

MATSUNAGA Takuya  Sapporo Medical University, School of Medicine, Assistant professor (70260768)

Co-Investigator(Kenkyū-buntansha) IYAMA Satoshi  Sapporo Medical University, School of Medicine, Instructor (50398319)
SATO Tsutomu  Sapporo Medical University, School of Medicine, Instructor (40404602)
Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsCord blood / CD34^+ cells / Artificial platelets
Research Abstract

We generated platelets (PLT) from cord blood (CB) CD34^+ cells employing a three-phase culture system. We first cultured 500 CB CD34^+ cells on telomerase gene-transduced human stromal cells (hTERT stroma) in serum-free medium supplemented with stem cell factor (SCF), Flt-3/Flk-2 ligand (FL) and thrombopoietin (TPO) for 14 days. We then transferred the cells to hTERT stroma and cultured for another 14 days with fresh medium containing interleukin-11 (IL-11) in addition to the original cytokine cocktail. Subsequently, we cultured the cells in a liquid culture medium containing SCF, FL, TPO and IL-11 for another 5 days to recover PLT fractions from the supernatant, which were then gel filtered to purify the PLT. The calculated yield of PLT from 1.0 unit CB (5x10^6 CD34+ cells) was 1.26 x10^<11> - 1.68 x10^<11> PLT. These numbers of PLT are equivalent to 2.5-3.4 units of random donor-derived PLT or 2/5-6/10 of a single apheresis PLT. The CB-derived PLT exhibited features quite similar to those from peripheral blood in morphology, as revealed by electron micrographs, and in function, as revealed by fibrinogen/ADP aggregation, with the appearance of P-selectin and activated glycoprotein IIb-IIIa antigens. Thus this culture system may be applicable for large scale generation of PLT for future clinical usage.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (34 results)

All 2008 2007 2006

All Journal Article (24 results) (of which Peer Reviewed: 13 results) Presentation (10 results)

  • [Journal Article] Ex vivo large scale generation of human platelets from cord blood CD34^+ cells2008

    • Author(s)
      Matsunaga T, et. al.
    • Journal Title

      Leukemia 22

      Pages: 353-360

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Aberrant crypt foci as precursors of the dysplasia-carcinoma sequence inpatients with ulcerative colitis2008

    • Author(s)
      Kukitsu T, Matsunaga T.
    • Journal Title

      Clin Cancer Res 14

      Pages: 48-54

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Ex vivo large-scale generarion of human red blood cells from cord blood CD34^+ cells by co-culturing with macrophages2008

    • Author(s)
      Fujimi A, Matsunaga T.
    • Journal Title

      Int J Hematol (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] A phase I trial of arterial infusion chemotherapy with gemcitabine and 5-fluorour acil for unresectable advanced pancreatic cancer after vascular supply distribution via super selecitive embolization2008

    • Author(s)
      Miyanishi K, Matsunaga T.
    • Journal Title

      Epn J Clin Oncol 38

      Pages: 268-274

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Resolution of liver cirrhosis using vitamin A coupled liposomes to deliver siRNA against a collaen-specific chaperone2008

    • Author(s)
      Sato Y, Matsunaga T.
    • Journal Title

      Nat Biotechnol 38

      Pages: 431-442

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Combination therapy of an anticancer drug with the FNIII14 peptide of fibronectin effectively overcomes cell adhesion-mediated drug resistance of acute myelogenous leukemia2008

    • Author(s)
      Matsunaga, T., Fukai, F.
    • Journal Title

      Leukemia 22

      Pages: 353-360

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Aberrant crypt foci as precursors of the dysplasia-carcinoma sequence in patients with ulcerative colitis2008

    • Author(s)
      Kukitsu, T., Matsunaga, T
    • Journal Title

      Clin Cancer Res 14

      Pages: 48-54

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Ex vivo large-scale generation of human red blood cells from cord blood CD34(+) cells by co-culturing with macrophages2008

    • Author(s)
      Fujimi, A., Matsunaga, T
    • Journal Title

      Int J Hematol (Epub ahead of print)

    • NAID

      10025158459

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] A Phase I trial of arterial infusion chemotherapy with gemcitabine and 5-fluorouracil for unresectable advanced pancreatic cancer after vascular supply distribution via super selective embolization.2008

    • Author(s)
      Miyanishi, K., Matsunaga. T
    • Journal Title

      Jpn J Clin Oncol. 38

      Pages: 268-274

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Resolution of liver cirrhosis using vitamin A coupled liposomes to deliver siRNA against a collagen-specific chaperone2008

    • Author(s)
      Sato, Y., Matsunaga, T
    • Journal Title

      Nat Biotechnol 26

      Pages: 431-442

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Essential role of protein kinase C {zeta} in transducing a motility signal induced by superoxide and a chemotactic peptide, fMLP2007

    • Author(s)
      Kuribayashi K, Matsunaga T.
    • Journal Title

      J Cell Biol 176

      Pages: 1049-1060

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Phase I study of S-1, docetaxel and cisplatin combination chemotherapy inpatients with unresectable metastatic gastric cancer2007

    • Author(s)
      Takayama T, Matsunaga T.
    • Journal Title

      Br J Cancer 97

      Pages: 851-856

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Combination therapy of alland-cancer drug with the FNInl4 peptide of fibronectin effectively overwmes cell adhesion mediated-drug resistance of acute myelogenous leukemia2007

    • Author(s)
      Matsunaga T, et. al.
    • Journal Title

      Leukemia (Epub ahead of print)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Essential role of protein kinase C {zeta} in transducing a motility signal induced by superoxide and a chemotactic peptide, fMLP.2007

    • Author(s)
      Kuribayashi, K., Matsunaga, T
    • Journal Title

      J Cell Biol 176

      Pages: 1049-1060

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Phase I study of S-1, docetaxel and cisplatin combination chemotherapy in patients with unresectable metastatic gastric cancer2007

    • Author(s)
      Takayama, T., Matsunaga, T
    • Journal Title

      Br J Cancer 97

      Pages: 851-856

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Essential role of protein kinase C {zeta} in transducing a motility signal induced by superoxide and a chemotactic peptide, fMLP.2007

    • Author(s)
      Kuribayashi K, et. al.
    • Journal Title

      J Cell Biol 176

      Pages: 1049-1060

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Phase I study of S-1, docetaxel and cisplatin combination chemotherapy in patients with uuresectable metastatic gastric cancer.2007

    • Author(s)
      Takayama T, et. al.
    • Journal Title

      Br J Cancer 97

      Pages: 851-856

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Combination therapy of an anti-cancer drug with the FNIII14 peptide of fibronectin effectively overcomes cell adhesion mediated-drug resistance of acute myelogenous leukemia.2007

    • Author(s)
      Matsunaga T, et. al.
    • Journal Title

      Leukemia (Epub ahead of print)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Hepatitis C virus core protein promotes proliferation of human hepatoma Huh-7 cells through enhancement of transforming growth factor-[alpha] expression via activation of NF-kB2006

    • Author(s)
      Sato Y, Matsunaga T.
    • Journal Title

      Gut 55

      Pages: 1801-1808

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Amelioration ofmurine dextran sulfate sodium-induced colitis by ex vivo extrace llular superoxide dismutase gene transfer2006

    • Author(s)
      Oku T, Matsunaga T.
    • Journal Title

      Inflamm Bowel Dis 12

      Pages: 630-640

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Hepatitis C virus core protein promotes proliferation of human hepatoma Huh-7 cells through enhancement of transforming growth factor-[alpha] expression via activation of NF-DB2006

    • Author(s)
      Sato, Y., Matsunaga, T
    • Journal Title

      Gut 55

      Pages: 1801-1808

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Amelioration of murine dextran sulfate sodium-induced colitis by ex vivo extracellular superoxide dismutase gene transfer2006

    • Author(s)
      Oku, T., Matsunaga, T
    • Journal Title

      Inflamm Bowel Dis 12

      Pages: 630-640

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Ex vivo large scale generation of human platelets from cord blood CD34* cells2006

    • Author(s)
      Matsunaga, T., Tanaka, I
    • Journal Title

      Stem Cells 24

      Pages: 2877-2887

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Ex vivo large-scale generation of human platelets from cord blood CD34^+ cells.2006

    • Author(s)
      Matsunaga T, Iyama S, Sato T, et al.
    • Journal Title

      Stem cells 24

      Pages: 2877-2887

    • Related Report
      2006 Annual Research Report
  • [Presentation] Normal restoration of rat cirrhosis employing siRNA/hap47 encapsulated in viramin A liposomes2007

    • Author(s)
      Sato Y, Matsunaga T.
    • Organizer
      The 17th APASL Conference
    • Place of Presentation
      Kyoto, Japan
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ex vivo large-scale generation of human red blood cells from cord blood CD34^+ cells by co-culturing with macrophages2007

    • Author(s)
      Matsunaga T, et. al.
    • Organizer
      The 36th Annual Meeting of the International Society for Experimental Hematology
    • Place of Presentation
      Hamburg,Genmany
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] A novel modality of cancer therapy targeting cancer stroma2007

    • Author(s)
      Niitsu Y, Matsunaga T.
    • Organizer
      The Sixth Japan-China joint conference for cancer research
    • Place of Presentation
      Toya,Japan
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Wnt3/RhoA/ROCK signaling pathway is involved in VLA6-dependent adhesion-mediated drug resistance of multiple myeloma2007

    • Author(s)
      Kobune M, Matsunaga T.
    • Organizer
      The 49th Annua lMeeting of American Societyof Hematology
    • Place of Presentation
      Atlanta,USA
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Normal restoration of rat liver cirrhosis employing siRNA/HAP47 encapsulated in vitamin A liposomes2007

    • Author(s)
      Sato, Y., Matsunaga, T
    • Organizer
      The 17th APASL Conference
    • Place of Presentation
      Kyoto, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Ex vivo large-scale generation of human red blood cells from cord blood CD34^+ cells by co-culturing with macrophages2007

    • Author(s)
      Matsunaga, T., Fujimi, A
    • Organizer
      The 36th Annual Meeting of the International Society for Experimental Hematology
    • Place of Presentation
      Hamburg, Germany
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] A novel modality of cancer therapy targeting cancer stroma2007

    • Author(s)
      Niitsu, Y., Matsunaga, T
    • Organizer
      The Sixth Japan-China joint conference for cancer research
    • Place of Presentation
      Toya, Japan
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Wnt3/RhoA/ROCK signaling pathway is involved in VLA6-dependent adhesion-mediated drug resistance of multiple myeloma2007

    • Author(s)
      Kobune, M., Matsunaga, T
    • Organizer
      The 49th Annual Meeting of American Society of Hematology
    • Place of Presentation
      Atlanta, USA
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Simultaneous Oral Session. Combination therapy of anti-cancer drug with FNIII14 peptide of fibronectin effectively overcome cell adhesion mediated-drug resistance of acute myelogenous leukemia2006

    • Author(s)
      Matsunaga T, et. al.
    • Organizer
      The 48th Annual Meeting of American Society of Hematology
    • Place of Presentation
      Orland, USA
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Combination therapy of anti-cancer drug with FNIII14 peptide of fibronectin effectively overcome cell adhesion mediated-drug resistance of acute myelogenous leukemia2006

    • Author(s)
      Matsunaga, T., Fukai, F
    • Organizer
      The 48th Annual Meeting of American Society of Hematology
    • Place of Presentation
      Orland, USA
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2006-04-01   Modified: 2016-04-21  

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