, Volume 44, Issue 1–2, pp 15–25 | Cite as

Characterisation of Tetraploid and Diploid Clones of Spodoptera frugiperda Cell Line

  • R.F. Jarman-Smith
  • C. Mannix
  • M. Al-Rubeai


We have isolated and characterised diploid and tetraploid clones from the normally heterologous Spodoptera frugiperda (Sf-9)cell line by dilution cloning technique. Tetraploid clones were found to have cell sizes in excess of 35% larger than that of the diploid clones. In contrast, the maximum cell numbers achieved in batch cultures of diploid clones were on average 185% higher than the tetraploid cell numbers. Growth rates and metabolic quotients during the exponential phase were similar for both clones. Tetraploid cells infected with wild-type and recombinant green fluorescent protein (GFP) baculovirus, resulted in more polyhedra or GFP product per cell. Importantly, the difference between the clones either completely diminished or reduced to 50% when the yield was assessed in terms of the amount of polyhedra or GFP per mL of medium, respectively. These results indicate that the existing heterogeneity in insect cell populations with respect to ploidy level, are correlated to cell growth and product yield.

AcNPV Cell size Flow cytometry GFP Insect cells Intracellular pH Spodoptera frugiperda 


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  1. Al-Rubeai M.A., Kloppinger M., Fertig G., Emery A.N. and Miltenburger H.G. 1992. Monitoring of Biosynthetic and Metabolic Activity in Animal Cell culture Using Flow Cytometric Methods. Animal Cell Technology: Developments, Processes and Products. Butterworth-Heinemann Ltd, London, pp. 301–307.Google Scholar
  2. American Type Culture Collection (ATCC), 1988. Catalogue of Cell Lines and Hybridomas 6th edn. American Type Culture Collection, 166 pp.Google Scholar
  3. Coulson J.M. and Richardson J.F. 1999. Chapter 4: Flow of fluids through granular beds and packed columns. Chemical Engineering Volume 2, ''Particle and Separation Processes'', 133 pp.Google Scholar
  4. Cowger N.L., O'Connor K.C., Hammond T.G., Lacks D.J. and Navar G.L. 1999. Characterization of bimodal cell death of insect cells in a rotating-wall vessel and shaker flask. Biotechnol. and Bioeng. 64(1): 14–26.Google Scholar
  5. Disney J.E. and McCarthy W.J. 1985. A modified technique for the improved characterisation of lepidopteran chromosomes from cell in culture. In Vitro Cell. Dev. Biol. 21(10): 563–568.Google Scholar
  6. Doverskog M., Bertram E., Ljunggren J., Ohman L., Sennerstam R. and Häggström L. 2000. Cell cycle progression in serum-free cultures of Sf-9 insect cells: modulation by conditioned medium factors and implications for proliferation and productivity. Biotechnol. Prog. 16: 837–846.Google Scholar
  7. Fertig G., Klöppinger M. and Miltenburger H.G. 1990. Cell cycle kinetics of insect cell cultures compared to mammalian cell cultures. Exp. Cell Res. 189: 208–212.Google Scholar
  8. Francki R.I.B., Fauquet C.M., Knudson D.L. and Brown F. 1991. Fifth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, Berlin.Google Scholar
  9. Ishaque A. and Al-Rubeai M. 1998. Use of intracellular pH and annexin-V flow cytometric assays to monitor apoptosis and its suppression by bcl-2 over expression in hybridoma cell culture. J. Immunol. Methods 221: 43–57.Google Scholar
  10. Jarman-Smith R.F., Mannix C. and Al-Rubeai M. 2002. Chromosomal instability in Spodoptera frugiperda Sf-9 cells. Biotechnol. Prog. 18: 623–628.Google Scholar
  11. Jones I. and Morikawa Y. 1996. Baculovirus vectors for expression in insect cells. Curr. Opin. Biotechnol. 7: 512–516.Google Scholar
  12. Kioukia A.K. 1994. Mammalian cell and insect cell-baculovirus systems and their responses to hydrodynamic and other stresses in agitated and sparged reactors. PhD thesis, University of Birmingham, UK.Google Scholar
  13. Kioukia N., Al-Rubeai M., Zhang Z., Emery A.N., Nienow A.W. and Thomas C.R. 1995. A study of infected and baculovirus infected Spodoptera frugiperda cells in T and spinner flasks. Biotechnol. Lett. 17(1): 7–12.Google Scholar
  14. Leelavatcharamas V. 1997. Growth, gamma interferon production and cell cycle in batch, continuous and perfusion culture of CHO cells. PhD thesis, University of Birmingham, UK.Google Scholar
  15. Lery X., Charpentier G. and Belloncik B. 1999. DNA content analysis of insect cells by flow cytometry. Cytotechnology 29: 103–113.Google Scholar
  16. Miller L. 1988. Baculovirus as gene expression vectors. Ann. Rev. Microbiol. 42: 177–199.Google Scholar
  17. O'Reilly D.R., Miller L.K. and Lucklow V. 1994. Baculovirus expression vectors: a laboratory manual. Freeman, New York.Google Scholar
  18. Swanson C.P. 1981. Cytogenetics: the chromosome in division, inheritance and evolution 2 edn. In: Merz T. and Young W.J. (ed.), Prentice Hall, Englewood Cliffs, NJ, 577 pp.Google Scholar
  19. Vaughn J.L., Goodwin R.H., Tompkins G.J. and McCawley P. 1977. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In vitro 13(4): 213–217.Google Scholar
  20. Vlak J.M. 1992. The biology of baculoviruses in vivo and in cultured insect cells. Workshop on Baculovirus and recombinant protein production processes, Interlaken, Switzerland, pp. 2–10.Google Scholar
  21. White M.J.D. 1973. Animal Cytology and Evolution, 3rd edn., Cambridge University Press, Cambridge, 961 pp.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • R.F. Jarman-Smith
  • C. Mannix
  • M. Al-Rubeai

There are no affiliations available

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