| Project/Area Number |
15200033
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | University of Tsukuba |
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Principal Investigator |
MIYOSHI Hirotoshi (2004) Doctoral Program in Advanced Biomedical Applications, Graduate School of Comprehensive Human Sciences, Assistant Professor, 大学院・人間総合科学研究科, 講師 (70292547)
大島 宣雄 (2003) 筑波大学, 基礎医学系, 教授 (50015971)
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| Co-Investigator(Kenkyū-buntansha) |
OOKAWA Keiko Doctoral Program in Advanced Biomedical Applications, Graduate School of Comprehensive Human Sciences, Assistant Professor, 大学院・人間総合科学研究科, 講師 (30251052)
三好 浩稔 筑波大学, 基礎医学系, 講師 (70292547)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥50,310,000 (Direct Cost: ¥38,700,000、Indirect Cost: ¥11,610,000)
Fiscal Year 2004: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000)
Fiscal Year 2003: ¥35,620,000 (Direct Cost: ¥27,400,000、Indirect Cost: ¥8,220,000)
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| Keywords | hematopoietic stem cell / stromal cell / fetal liver cell / artificial bone marrow / growth / differentiation / three-dimensional culture / porous scaffold / tissue engineering / 胎子肝臓細胞 / 共培養 / シグナル分子 |
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| Research Abstract |
To develop a tissue-engineered artificial immune system, three-dimensional (3-D) coculture of hematopoietic cells and stromal cells were performed using porous polyvinyl formal (PVF) resin as a substrate material. In this series of culture experiments, mouse fetal liver cells (FLCs) were used as a hematopoietic cell source, and effects of stromal cell lines and 3-D cryopreservation of these cell lines on differentiation and expansion of hematopoietic stem cells were investigated.
When FLCs were cocultured with DAS 104-8 stromal cell line, 3-D culture for 2 weeks resulted in 5-fold expansion in hematopoietic progenitor cells (HPCs). To expand HPCs more conveniently, 3-D cryopreserved stromal cells which were made by cryopreserving stromal cells immobilized within the PVF resin cubes, were applied to the cultures of hematopoietic cells. After 2 weeks of coculture experiments, HPCs were expanded over a 15-fold, and these efficiencies were far better than the cultures using stromal cells without cryopreservation. Similar results were obtained when another stromal cell line (DAS 104-4) was used. Moreover, in the culture experiments using DAS 104-8 of which growth was suppressed by mitomycin C treatment, higher (15-fold) expansion of HPCs was also achieved than those using untreated DAS 104-8. Therefore, it was clarified that efficiencies of HPC expansion were improved by applying 3-D cryopreserved stromal cell lines mainly due to the suppressed growth of these cell lines.
To facilitate specific differentiation of HPCs into B cell lineage, FLCs were cocultured with DAS 104-8 cell line in the presence of interleukin-7. However, numbers of B cells as well as HPCs in the PVF cubes decreased rapidly with elapse of culture period. Therefore, further investigations concerning differentiation into B cells will be required.
In conclusion, efficient expansion system of HPCs was established by using 3-D cryopreserved stromal cells.
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