Three dimensional analyses of functions of cultured cells

为了对培养的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，发展了一种昆虫细胞的三维培养方法，该方法已应用于哺乳动物细胞，以研究其结构和细胞群落，但尚未在昆虫细胞中开发。对于新的细胞系，我们建立了两个功能细胞系：来自鞘翅目脂肪体组织的XP-1和来自鳞翅目脂肪体组织的TH-1。对这些细胞系的鉴定表明，XP-1对保幼激素和20-羟基蜕皮激素有不同程度的反应，而TH-1对AcNPV敏感。我们还检测了来自白纹伊蚊全组织的AEL-2和来自油菜Mamestra brassicae血细胞的MABR-93等其他细胞系的功能。对这些细胞的功能分析表明，AELL-2细胞产生乙酰胆碱酯酶受保幼激素、20-羟基蜕皮激素和细胞群结构的影响，MaBR-93细胞分泌的耐热生长因子的相对分子质量小于5000。在这些细胞系中，AEL-2细胞成功地保持在三维培养中，尽管制备了与昆虫细胞严重相关的胶原网。在胶原网中培养的AEL-2细胞与在液体中二维培养的AEL-2细胞表现不同，乙酰胆碱酯酶的产生和细胞伸展的形态也不同。因此，昆虫细胞的三维培养可能诱导其在二维培养环境中潜伏的功能。

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.