2004 Fiscal Year Final Research Report Summary
Low-cost ex-vivo expansion system for hematopoietic progenitor cells employing a membrane-separated coculture system of hematopoietic cells and stromal cell line.
| Project/Area Number |
14380400
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | National Hospital Organization, Osaka Minami Medical Center, Clinical Research Institute (2003-2004)
Osaka University (2002) |
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Principal Investigator |
SOMA Toshihiro National Hospital Organization, Osaka Minami Medical Center, Clinical Research Institute, Department of Clinical Laboratory, Director, 大阪南医療センター・臨床検査部, 部長 (40273619)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAGI Mutsumi Hokkaido University, Graduate School b of Engineering, Professor, 工学研究科・分子化学専攻, 教授 (20263212)
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| Project Period (FY) |
2002 – 2004
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| Keywords | hematopoietic cells / bone marrow transplantation / co-culture / porous membrane / reactor / cord blood / three-dimensional culture / nonwoven fabrics |
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| Research Abstract |
The effect of pore diameter of porous membrane on the content of progenitor cells during the membrane-separated coculture of murine bone marrow hematopoietic cells and a murine stromal cell line, in which stromal cells adhered onto the lower surface of the membrane and hematopoietic cells were incubated on the upper surface of the membrane, was investigated in order to design a membrane bioreactor for ex vivo expansion of hematopoietic primitive cells employing exogeneic stromal cells. The CFU-Mix content at 1 week in the membrane-separated coculture increased as the pore diameter of the membrane decreased from 12.0 to 0.4 μm. However, the decrease in pore diameter from 0.4 to 0.1 μm did not affect the CFU-Mix content. Observation of stromal cells that adhered on the lower surface of the porous membranes (0.4 and 3.0 μm pore diameters) under a confocal scanning microscope after staining with rhodamine phalloidin suggested that stromal cells reached the membrane's upper surface through the pore with a diameter larger than 3.0 μm. Consequently, membrane having small (≦0.4 μm) pores that stromal cells could not pass through prevented the direct contact between stromal and hematopoietic cells, which resulted in a higher content of hematopoietic progenitor cells (CFU-Mix) during the membrane-separated coculture.
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