2003 Fiscal Year Final Research Report Summary
The development of culture system for platelet substitute production using human hematopoietic stem cells and megakaryocytic cell line
| Project/Area Number |
14571000
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | Saitama Medical University (2003)
Tokyo Medical University (2002) |
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Principal Investigator |
IKEBUCHI Kenji Saitama Medical University, Department of Transfusion and Cell Therapy, Professor, 医学部, 教授 (20175194)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Platelet / Substitute / Megakaryocytic line / Bone marrow stroma / human cord blood / CD34+ cell / SCIDマウス |
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| Research Abstract |
Human cord blood CD34+ cells produced approximately 1000-fold hematopoietic cells and 100-fold CD34+ cells in the culture system consisting of TPO, FL, SCF and human bone marrow-derived stroma cells. The function of stroma cells required cell-cell direct contact and this capability was partially preserved after chemical fixation.
Cord blood C34+ cells differentiated to megakaryocytic cells after culture in the presence of TPO and SCF, however, massive expansion was not obtained. In addition, the terminal maturation such as proplatelet formation, was not observed. Human megakaryocytic cell line(Meg-J) cells produced platelet-like particles in the culture. The addition of TPA or protein kinase inhibitor promoted cell maturation, together cell apoptosis. Platelet-particles consisted of cellular fragment and apoptotic bodies.
In order to permanently use hematopoietic cytokines in culture, the method to coat cytokines on culture device surface area was examined. Photoreactive gelatin and erythropoietin(EPO) was fixed on the plastic surface by use of UV irradiation. Epo-dependent cell line grew on EPO-fixed surface, but did not grow on negative area, indicating that this coating method was applicable.
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Research Products
(8 results)
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[Publications] Hirayama F, Yamaguchi M, Yano M, Yasui K, Horie Y, Matsumoto K, Nagao N, Ikebuchi K, Azuma H, Ikeda H, Tani Y: "Spontaneous and rapid reexpression of functional CXCR4 by human steady-state peripheral blood CD34+ cells"Int J Hematol. 78(1). 48-55 (2003)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kawano Y, Kobune M, Yamaguchi M, Nakamura K, Ito Y, Sasaki K, Takahashi S, Nakamura T, Chiba H, Sato T, Matsunaga T, Azuma H, Ikebuchi K, Ikeda H, Kato J, Niitsu Y, Hamada H: "Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human catalytic subunit(hTERT)-transfected human stromal cells."Blood. 101(2). 532-540 (2003)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kawano Y, Kobune M, Yamaguchi M, Nakamura K, Ito Y, Sasaki K, Takahashi S, Nakamura T, Chiba H, Sato T, Matsunaga T, Azuma H, Ikebuchi K, Ikeda H, Kato J, Niitsu Y, Hamada H: "Ex vivo expansion of human umbilical cord hematopoietic progenitor cells using a coculture system with human telomerase catalytic subunit(hTERT)-transfected human stromal cells."Blood. 101(2). 532-540 (2003)
Description
「研究成果報告書概要(欧文)」より
