| Project/Area Number |
14580809
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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 |
Biomedical engineering/Biological material science
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| Research Institution | University of Tsukuba |
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Principal Investigator |
MIYOSHI Hirotoshi University of Tsukuba, Department of Biomedical Engineering, Institute of Basic Medical Sciences, Assistant Professor, 基礎医学系, 講師 (70292547)
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| Co-Investigator(Kenkyū-buntansha) |
OOKAWA Keiko University of Tsukuba, Department of Biomedical Engineering, Institute of Basic Medical Sciences, Assistant Professor, 基礎医学系, 講師 (30251052)
OHSHIMA Norio University of Tsukuba, Department of Biomedical Engineering, Institute of Basic Medical Sciences, Professor, 基礎医学系, 教授 (50015971)
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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 |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | hepatocyte / fetal liver cell / signaling molecule / three-dimensional culture / perfusion culture / porous scaffold / packed-bed reactor / tissue engineering / ブタ胎仔肝臓細胞 |
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| Research Abstract |
To develop a tissue-engineered bioartificial liver, long-term three-dimensional (3-D) cultures of fetal liver cells (FLCs) utilizing porous polymer as a 3-D scaffold were performed, and effects of signaling molecules on proliferation and maturation of FLCs were investigated. To examine the effects of medium perfusion rate on maintenance of cell numbers and metabolic functions, perfusion cultures of FLCs using a packed-bed type reactor system were also performed.
In the perfusion culture experiments using a packed-bed type reactor, cell numbers and hepatic functions of pig FLCs were stably maintained over one month, although those of mouse FLCs rapidly decreased. Thus, pig FLCs were considered to be applicable as a cell source of bioartificial livers.
To examine the effects of medium perfusion on functions and morphology of cultured cells more closely, perfusion cultures of adult rat hepatocytes were performed using a parallel-plate flow chamber. In this series of experiments, it was clarified that medium perfusion induced 3-D reconstruction of cocultured hepatocytes with nonparenchymal cells, and morphology of these cells were similar to that of in vivo liver.
With respect to the signaling molecules, effects of oncostatin M, epidermal growth factor and dimethyl sulfoxide on proliferation and metabolic functions of 3-D cultured FLCs were investigated. These factors strongly induced the maturation of FLCs under the 3-D culture condition. Moreover, when the cultured cells were temporally stimulated by controlling supply of these factors, the maturation was facilitated.
From these facts, the 3-D culture method of FLCs in this study was considered to be applicable to developing tissue-engineered bioartificial livers.
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