title: Cell Culture for Production of Insecticidal Viruses.
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.
authors: School of Chemistry & Molecular Biosciences, The University of Queensland, Brisbane, QLD, 4072, Australia. steven.reid@uq.edu.au.
authors: Steven
authors: S
authors: Reid
authors: Patheon Biologics Australia Pty Ltd, 37 Kent Street, Woolloongabba, QLD, 4156, Australia.
authors: Leslie C L
authors: LC
authors: Chan
authors: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, 4072, Australia.
authors: Leila
authors: L
authors: Matindoost
authors: Laboratory for Evolution and Development, Department of Biochemistry, University of Otago, Dunedin, New Zealand.
authors: Charlotte
authors: C
authors: Pushparajan
authors: Chemical & Process Engineering Department, University of Canterbury, Canterbury, New Zealand.
authors: Gabriel
authors: G
authors: Visnovsky
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques; D061046:Biological Control Agents; D019149:Bioreactors; D018929:Cell Culture Techniques; D002460:Cell Line; D003470:Culture Media; D005285:Fermentation; D007313:Insecta; D007306:Insecticides; D014780:Viruses
chemical_list: D061046:Biological Control Agents; D003470:Culture Media; D007306:Insecticides
keywords: Adherent culture; Bioreactors; Insect cell technology; Insecticidal viruses; Suspension culture