| Project/Area Number |
63890011
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
広領域
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SASAKI Ryuzo Kyoto University, Agriculture, Professor, 農学部, 教授 (60077378)
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| Co-Investigator(Kenkyū-buntansha) |
UEDA Masatsugu Snow Brand Milk Products Co., Ltd., Research Institute of Life Science, Chief Sc, 生物科学研究所, 課長
SHIRAI Yoshihito Kyoto University, Engineering, Assistant, 工学部, 助手 (50175395)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 1989: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1988: ¥8,500,000 (Direct Cost: ¥8,500,000)
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| Keywords | Cultured animal cells / Immobilization of cells / Alginate gels / Microcarrier / Recombinant erythropoietin / Erythropoietic factor / Anchorage-dependent cells / マイクロキャリア- / 組み換え型エリスロポエチン |
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| Research Abstract |
Recombinant glycoproteins have to be produced by animal cells, since procaryotic cells do not glycosylated. So far these proteins have been produced by anchorage-dependent cells and therefore, development of the method that makes it possible to proliferate these cells in a high density is important for production of these valuable proteins in a high efficiency. In this project, we attempted to establish the method in which the anchorage-dependent cells producing the erythropoietic factor were immobilized in alginate gels. Immobilization would protect the cells from harmful physical forces and alginate gels may provide the wide space for growth of the cells.
Erythropoietin (EPO), a glycoprotein that is a physiological stimulator of erythrocyte production, was produced continuously for more than 32 days by three kinds of anchorage-dependent animal cells immobilized in alginate gel particles. Gelation caused by divalent cations added to an alginate solution containing cells resulted in the formation of clear vacant spaces (referred to here as channels). Each channels originated from a cell and extended towards the center of the gel particle. The animal cells grow well in the channels but proliferated little outside the channels. The cell concentration in the gel particles reached more than 1 x 10 cells/g gel. The alginate immobilization method was useful for high-concentration cultivation of the anchorage-dependent cells.
Baby hamster kidney (BHK) cells engineered to produce recombinant human EPO were cultured at high density on microcarriers entrapped by calcium alginate gel particles. In this system, the BHK cells proliferated not only on the microcarriers but also in vacant spaces in the alginate gel particles. These spaces contributed to high-density cultivation of the cells and a high productivity of EPO.
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