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Establishment, growth kinetics, and susceptibility to AcMNPV of heat tolerant lepidopteran cell lines


Lepidopteran heat-tolerant (ht) cell lines have been obtained with sf-9, sf-21 and several Bombyx cells. They have a distinct karyotype, membrane lipid composition, morphology and growth kinetics from the parental cell lines. In this paper, we report the development of ht cell lines from other insect species and examination of their growth characteristics and virus susceptibility. Adaptation of cell lines sf-9, BTI-TN-5B1-4 (High5) and BTI-TN-MG1 (MG1) to 33°C and 35°C was carried out by shifting the culture temperature between 28°C and higher temperatures by a gradual stepwise increase in temperature. The process of adaption to a higher culture temperature was accomplished over a period of 2 months. The cell lines with the temperature adaption were designated as sf9-ht33, sf9-ht35, High5-ht33, High5-ht35, MG1-ht33, MG1-ht35. These cell lines have been subcultured over 70 passages. Adaption to high temperatures was confirmed by a constant population doubling time with individual cell lines. The population doubling time of heat adapted cell lines were 1–4 h less than these of parental cell lines. Cell shapes did not show obvious change, however, the cell size of sf9-ht cells was enlarged and those of High5 and MG1 ht cells were reduced after heat adaption. When the cell lines were infected with Autographa californica nuclear polyhedrosis virus (AcMNPV) at 28°C, 33°C, 35°C and 37°C, production of budded virus and occlusion bodies in each cell line was optimum at its own adapted temperature.

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  1. Carretero M T, Carmona M J, Diez J L. 1991. Thermotolerance and heat shock proteins in Chironomus. J Insect Physiol, 37: 239–246.

    Article  CAS  Google Scholar 

  2. Carvalho M G, Freitas M S. 1988. Effect of continues heat stress on cell growth and protein synthesis in Aedes albopictus. J Cell Physiol, 137: 455–461.

    Article  CAS  PubMed  Google Scholar 

  3. Earley E M. 1975. Chromosomal preparation from monolayer cell cultures. Methods Cell Sci, 1: 31–35.

    CAS  Google Scholar 

  4. Fittinghoff C M, Riddiford L M. 1990. Heat sensitivity and protein synthesis during heat-shock in the tobacco hornworm, Manduca sexta. J Comp Physiol B, 160: 349–356.

    Article  CAS  PubMed  Google Scholar 

  5. Gerbal M, Fournier P, Barry P, et al. 2000. Adaptation of an insect cell line of Spodoptera frugiperda to grow at 37°C: characterization of an endodiploid clone. In Vitro Cell Dev Biol-Anim, 36: 117–124.

    Article  CAS  PubMed  Google Scholar 

  6. Grace T D C. 1962. Establishment of four strains of cells from insect tissues grown in vitro. Nature, 195: 788–789.

    Article  CAS  PubMed  Google Scholar 

  7. Grace T D C. 1967. Insect cell culture and virus research. In Vitro Cell Dev Biol — plant, 3: 104–117.

    Article  Google Scholar 

  8. Granados R R, Li G, Blissard G W. 2007. Insect cell culture and biotechnology. Virol Sin, 22: 83–93.

    Article  CAS  Google Scholar 

  9. Granados R R, Li G, Derksen A C G, et al. 1994. A new insect cell line from Trichoplusia ni (BTI-Tn-5B1-4) susceptible to Trichoplusia ni singly enveloped nuclear polyhedrosis virus. J Invertebr Pathol, 64: 260–266.

    Article  Google Scholar 

  10. Hayflick L. 1973. Theory of population increase by subcultivation. In: Tissue Culture Methods and Application (Kruse P E & Patterrson M K. eds.), Pulishing place: Academic Press, New York. p222–223.

    Google Scholar 

  11. Imanishi S, Cho E S, Tomita S. 1999. Novel Bombyx mori cell lines cultivable at 37°C. Appl Entomol Zool, 34: 259–266.

    Google Scholar 

  12. Li G X, Hashimoto Y, Granados R R. 2003. Growth characteristics and expression of recombinant protein by new cell clones derived from Trichoplusia ni (BTI-Tn-5B1-4) High Five cells. Bioprocessing, 2: 34–37.

    Google Scholar 

  13. Lindquist S, Graig E A. 1988. The heat shock proteins. Annu Rev Genet, 22: 631–677.

    Article  CAS  PubMed  Google Scholar 

  14. Lynn D E. 2001. Effects of temperature on the susceptibility of insect cells to infection by baculovirus. Methods Cell Sci, 23: 221–225.

    Article  CAS  PubMed  Google Scholar 

  15. Meng M J, Li T L, Li C Y, et al. 2008. A suspended cell lines from Trichoplusia ni (Lepidoptera): Characterization and expression of recombinant proteins. Insect Sci, 15:423–428.

    Article  CAS  Google Scholar 

  16. Mitchell H K, Moller G, Petersen N S. 1979. Specific protection from phenocopy induction by heat shock. Dev Genet, 1:181–192.

    Article  CAS  Google Scholar 

  17. Ritossa F M. 1962. A new puffing pattern induced by heat shock and DNP in Drosophila. Experimentia, 18: 571–573.

    Article  CAS  Google Scholar 

  18. Wang P, Granados R R, Shuler M L. 1992. Studies on serum-free culture of insect cells for virus propagation and recombinant protein production. J Invertebr Pathol, 59: 46–53.

    Article  CAS  Google Scholar 

  19. Wang P, Toung R, Granados R R. 1999. The establishment of new cell lines from Pseudaletia unipuncta with differerntial responses to baculovirus infection. In Vitro Cell Dev Biol Animal, 35: 333–338.

    Article  CAS  Google Scholar 

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Correspondence to Guo-xun Li.

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Foundation items: Chinese National Basic Research Program (973) 2009CB118900; Chinese National Science Foundation Project (30771451); Boyce Thompson Institute for Plant Research Project (BTI-QAU 1-23-2007).

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Wu, Yl., Jiang, L., Hashimoto, Y. et al. Establishment, growth kinetics, and susceptibility to AcMNPV of heat tolerant lepidopteran cell lines. Virol. Sin. 26, 198–205 (2011).

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Key words

  • Heat-tolerant cell lines
  • Susceptibility of virus
  • Production of virus
  • Growth curves
  • Insect cell lines