Three dimensional analyses of functions of cultured cells

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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.