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Three dimensional analyses of functions of cultured cells

培养细胞功能三维分析

基本信息

批准号：
09460025
负责人：
IWABUCHI Kikuo
金额：
$ 7.87万
依托单位：
Tokyo University of Agriculture & Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460025/
关键词：
three-dimentional culture insect cell line acetylcholinesterase mosguiTo mosquito

项目摘要

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.

为了对昆虫细胞进行三维分析，我们建立了新的功能细胞系作为材料，并发展了昆虫细胞三维培养方法，该方法已用于哺乳动物细胞，以研究其结构和细胞群落，但尚未在昆虫细胞中发展。在新细胞系方面，我们建立了两个功能细胞系：鞘翅目脂肪体组织来源的XP-1和鳞翅目脂肪体组织来源的TH-1。这些细胞系的特性表明，XP-1是响应保幼激素和20-羟基蜕皮激素以不同的方式，和TH-1是敏感的AcNPV。我们还研究了其他细胞系的功能，AeAl-2来自白纹伊蚊的整个组织和MaBr-93来自甘蓝夜蛾的血细胞。这些细胞的功能分析表明，AeAl-2的乙酰胆碱酯酶的生产受到保幼激素，20-羟基蜕皮激素，和细胞群体的建设，MaBr-93细胞分泌的分子量小于5000的热稳定生长因子。在这些细胞系中，AeAl-2细胞成功地维持在三维培养中，尽管用昆虫细胞制备胶原晶格严重。AeAl-2细胞在胶原网格中培养的行为与二维液体培养基中培养的行为不同，乙酰胆碱酯酶的产生和细胞延伸的形态也不同。因此，昆虫细胞的三维培养可以诱导其在二维培养环境中潜伏的功能。