Lepidopteran cell lines after long-term culture in alternative media: Comparison of growth rates and baculovirus replication
- Dwight E. Lynn
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Three insect cell lines, IPLB-LdFB and IPLB-LdEIta from gypsy moth fat body and embryos and UFL-AG-286 from velvetbean caterpillar embryos, have been concurrently maintained for 1 to 12 yr on two media formulations, modified TC-100 containing 9% fetal bovine serum and Ex-cell 400, a commercial serum-free medium (SFM). Cells grown in each medium were tested for susceptibility to and productivity of various multiply embedded nucleopolyhedroviruses. The three lines chosen for these experiments fall into three categories of relative growth in SFM versus TC-100: LdFB cells grew similarly in each medium, LdEIta grew better in Ex-Cell than in TC-100, and AG-286 grew better in TC-100 than in Ex-Cell. The susceptibility of cells to infection also varies, although without any apparent correlation to which medium was best for supporting growth. Endpoint assays suggested that LdFB cells grown in serum-containing medium are more susceptible to virus infection than their SFM counterparts, while the opposite is true for LdEIta cells. Production of virus, based on numbers of occlusion bodies, showed fewer differences with only AcMNPV production with AG-286 in TC-100 being statistically higher than production of the same virus in Ex-cell 400. These studies suggest that long-term passage in alternative media may impact the ability of cells to support virus infection and replication, but the effects on each cell line and virus system need to be determined.
- Gardiner, G. R.; Stockdale, H. Two tissue culture media for production of lepidopteran cells and nuclear polyhedrosis viruses. J. Invertebr. Pathol. 25:363–370; 1975. CrossRef
- Goodwin, R. H.; Adams, J. R. Nutrient factors influencing viral replication in serum-free insect cell line culture. In Kurstak, E.; Maramorosch, K.; Dübendorfer, A., ed. Invertebrate systems in vitro. Amsterdam: Elsevier 1980:493–509.
- Grace, T. D. C. Establishment of four strains of cells from insect tissue grown in vitro. Nature 195:788–789; 1962. CrossRef
- Guzo, D.; Dougherty, E. M.; Braun, S. K.; Lynn, D. E.; Weiner, R. M. Production of a cellular macromolecular synthesis inhibition factor(s) in gypsy moth cells infected with the Autographa californica nuclear polyhedrosis virus. J. Invertebr. Pathol. 57:413–425; 1991. CrossRef
- Lynn, D. E. A BASIC computer program for analyzing endpoint assays. Biotechniques 12:880–881; 1992.
- Lynn, D. E. Available lepidopteran insect cell lines. In: Murhammer, D. W. ed. Methods in molecular biology: baculovirus protocols. Totowa, NJ: Humana Press; in press.
- Lynn, D. E. Comparative susceptibilities of twelve insect cell lines to infection by three baculoviruses. J. Invertebr. Pathol. 82:129–131; 2003. CrossRef
- Lynn, D. E. Comparison of cell line maintenance procedures on insect cells used for producing baculoviruses. In Vitro Cell. Dev. Biol. 35A:248–251; 1999.
- Lynn, D. E. Effects of temperature on the susceptibility of insect cells to infection by baculoviruses. Methods Cell Sci. 23:221–225; 2002. CrossRef
- Lynn, D. E. Methods for the development of cell lines from insects. Journal of Tissue Culture Methods 12:23–29; 1989. CrossRef
- Lynn, D. E.; Dougherty, E. M.; McClintock, J. T.; Loeb, M. Development of cell lines from various tissues of Lepidoptera. In: Kuroda, Y.; Kurstak, E.; Maramorosch, K., eds. Invertebrate and fish tissue culture. Tokyo: Japan Scientific Societies Press; 1988:239–242.
- Lynn, D. E.; Shapiro, M.; Dougherty, E. M. Selection and screening of clonal isolates of the abington strain of gypsy moth nuclear polyhedrosis virus. J. Invertebr. Pathol. 62:191–195; 1993. CrossRef
- Sieburth, P. J.; Maruniak, J. E. Growth characteristics of a continuous cell line from the velvetbean caterpillar, Anticarsa gemmatalis Hübner (Lepidoptera: Noctuidae). In Vitro Cellular and Developmental Biology 24:195–198; 1988. CrossRef
- Summers, M. D.; Smith, G. E. A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin No. 155556; 1987.
- Lepidopteran cell lines after long-term culture in alternative media: Comparison of growth rates and baculovirus replication
In Vitro Cellular & Developmental Biology - Animal
Volume 42, Issue 5-6 , pp 149-152
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- insect cell lines
- serum-free medium
- virus productivity
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- Dwight E. Lynn (1)
- Author Affiliations
- 1. Insect Biocontrol Lab, BARC West, USDA/ARS, Building 011A, Room 214, 10300 Baltimore Boulevard, 20705-2350, Beltsville, Maryland