Evaluation of concentrated milk whey as a supplement for SF9 Spodoptera frugiperda cells in culture
Fabiana R. X. Batista
Carlos A. Pereira
Ronaldo Z. Mendonça
Angela M. Moraes*
Financial support: 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 Butantan Foundation (São Paulo, Brazil). C.A. Pereira is recipient of a fellowship from the Conselho Nacioanl de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, Brazil).
Keywords: baculovirus, concentrated milk whey, Grace's medium, insect cells, media formulation, Spodoptera frugiperda Sf9.
Insect cell culture has become increasingly useful for the production of heterologous proteins as well as of baculovirus polyhedra, and several different culture media formulations can be employed for this purpose. The goal of this work was to assess the potential of lyophylized milk whey ultra filtration retentate (MWR) when associated to yeast extract (YE), glucose and Pluronic F68 (PF68) to partially replace fetal calf serum (FBS) in Spodoptera frugiperda Sf9 cells culture in Grace's medium, aiming AgMNPV baculovirus production. Batch cultivation results showed that the yeast extract and the milk whey concentrate effectively increased cell concentration to about half the level commonly verified for the Sf900II serum-free medium. A 4-fold increase in viable cell concentration was achieved when employing 5% (w/v) MWR, 8 g/L YE, 1% (v/v) of FBS and 2.7 g.l-1 of glucose, resulting in 1.6 x 107 polyhedra.ml-1 after infection with baculovirus.
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