| 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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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Platelet / Substitute / Megakaryocytic line / Bone marrow stroma / human cord blood / CD34+ cell / SCIDマウス / 造血幹細胞 / 増幅 / ストロマ細胞 / 化学固定 / 巨核球 |
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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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