Abstract
An animal protein-free medium was developed for Drosophila melanogaster S2 (S2AcGPV2) cells genetically modified to produce the rabies virus G glycoprotein (GPV). IPL-41, used as a basal medium, was supplemented with yeastolate, carbohydrates, amino acids and lipids aiming initially to reduce and further to eliminate the need of fetal bovine serum. The S2AcGPV2 cells were fully capable of growing in serum-free supplemented IPL-41 medium containing 6 g L−1 yeastolate ultrafiltrate, 10 g L−1 glucose, 3.5 g L−1 glutamine, 0.5 g L−1 fructose, 2 g L−1 lactose, 0.6 g L−1 tyrosine, 1.48 g L−1 methionine and 1% (v/v) lipid emulsion, reaching 19 × 106 cells mL−1. Maximum specific growth rate and cell productivity were 0.025 h−1 and 0.57 × 105 cells mL−1 h−1, respectively. Glucose and lactose were consumed during cell culture, but not fructose. Lactate concentration generally decreased during cell culture, while ammonium concentration reached 167 mg L−1, however, without noticeable deleterious effects on cell growth. GPV concentration values achieved were, however, modest in the proposed medium formulation.
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Acknowledgments
The authors acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, Brazil), Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP grant no. 02/09482-3, São Paulo, Brazil) and Instituto Butantan (São Paulo, Brazil) for the financial support. The authors wish to acknowledge Prof Dr. Cláudio Alberto Torres Suazo (Federal University of São Carlos, Brazil) and team for amino acids analysis. We are also grateful to Prof Dr. Andreas Karoly Gombert and Saul Nitsche Rocha from University of São Paulo for sugars measurements.
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Â. M. Moraes and C. A. Pereira are recipient of CNPq fellowships.
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Batista, F.R.X., Pereira, C.A., Mendonça, R.Z. et al. Formulation of a protein-free medium based on IPL-41 for the sustained growth of Drosophila melanogaster S2 cells. Cytotechnology 57, 11–22 (2008). https://doi.org/10.1007/s10616-008-9153-0
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DOI: https://doi.org/10.1007/s10616-008-9153-0