| Project/Area Number |
09460025
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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 |
植物保護
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| Research Institution | Tokyo University of Agriculture & Technology |
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Principal Investigator |
IWABUCHI Kikuo Tokyo University of Agriculture and Technology, Faculty of Agriculture, Associate Professor, 農学部, 助教授 (00203399)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAOKA Tsuyoshi Tokyo University of Agriculture and Technology, Faculty of Agriculture, Assistant, 農学部, 助手 (10238339)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥11,100,000 (Direct Cost: ¥11,100,000)
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| Keywords | three-dimentional culture / insect cell line / acetylcholinesterase / mosguiTo / mosquito |
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| Research Abstract |
To analyze cultured insect cells three-dimentionally, we established new functional cell lines as materials and developed a three-dimentional culture method for insect cells, which has been used in mammalian cells in order to examine their structure and cellular communities, but has not been developed in insect cells. As for new cell line, we established two functional cell lines as follows : XP-1 derived from coleopteran fat body tissues and TH-1 from lepidopteran fat body issues. Characterization of these cell lines showed that XP-1 was responsive to juvenile hormone and 20-hydroxyecdysone in different ways, and TH-1 was susceptible to AcNPV. We also examined the function of other cell lines, AeAl-2 derived from whole tissues of Aedes albopictus and MaBr-93 derived from hemocytes of Mamestra brassicae. The analyses of functions of these cells showed that the production of acetylcholinesterase by AeAl-2 was affected by juvenile hormone, 20-hydroxyecdysone, and construction of cell population, and that MaBr-93 cells secrete a thermostable growth factor of molecular weight less than 5000. Among these cell lines, AeAl-2 cells were successfully maintained in a three-dimentional culture, inspite of preparation of collagen lattices severe with insect cells. AeAl-2 cells cultured in the collagen lattice behaved differently from that cultured two-dimentionally in liquid medium, and the production of acetylcholinesterase and morphology of cellular extensions were also different. Thus, three-dimentional culture of insect cells may induce their functions which are latent in two-dimentional culture environment.
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