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Cell Culture for Production of Insecticidal Viruses

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Microbial-Based Biopesticides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1477))

Abstract

While large-scale culture of insect cells will need to be conducted using bioreactors up to 10,000 l scale, many of the main challenges for cell culture-based production of insecticidal viruses can be studied using small-scale (20–500 ml) shaker/spinner flasks, either in free suspension or using microcarrier-based systems. These challenges still relate to the development of appropriate cell lines, stability of virus strains in culture, enhancing virus yields per cell, and the development of serum-free media and feeds for the desired production systems. Hence this chapter presents mainly the methods required to work with and analyze effectively insect cell systems using small-scale cultures. Outlined are procedures for quantifying cells and virus and for establishing frozen cells and virus stocks. The approach for maintaining cell cultures and the multiplicity of infection (MOI) and time of infection (TOI) parameters that should be considered for conducting infections are discussed.

The methods described relate, in particular, to the suspension culture of Helicoverpa zea and Spodoptera frugiperda cell lines to produce the baculoviruses Helicoverpa armigera nucleopolyhedrovirus, HearNPV, and Anticarsia gemmatalis multicapsid nucleopolyhedrovirus, AgMNPV, respectively, and the production of the nonoccluded Oryctes nudivirus, OrNV, using an adherent coleopteran cell line.

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Author information

Authors and Affiliations

  1. School of Chemistry & Molecular Biosciences, The University of Queensland, Brisbane, QLD, 4072, Australia

    Steven Reid

  2. Patheon Biologics Australia Pty Ltd, 37 Kent Street, Woolloongabba, QLD, 4156, Australia

    Leslie C. L. Chan

  3. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, 4072, Australia

    Leila Matindoost

  4. Laboratory for Evolution and Development, Department of Biochemistry, University of Otago, Dunedin, New Zealand

    Charlotte Pushparajan

  5. Chemical & Process Engineering Department, University of Canterbury, Canterbury, New Zealand

    Gabriel Visnovsky

Authors
  1. Steven Reid
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  2. Leslie C. L. Chan
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  3. Leila Matindoost
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  4. Charlotte Pushparajan
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  5. Gabriel Visnovsky
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Corresponding author

Correspondence to Steven Reid .

Editor information

Editors and Affiliations

  1. Bio-Protection Research Centre, Lincoln University, Lincoln, New Zealand

    Travis R. Glare

  2. Bio-Protection Research Centre, Lincoln University, Lincoln, New Zealand

    Maria E. Moran-Diez

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Reid, S., Chan, L.C.L., Matindoost, L., Pushparajan, C., Visnovsky, G. (2016). Cell Culture for Production of Insecticidal Viruses. In: Glare, T., Moran-Diez, M. (eds) Microbial-Based Biopesticides. Methods in Molecular Biology, vol 1477. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6367-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-6367-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6365-2

  • Online ISBN: 978-1-4939-6367-6

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