Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media
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An animal protein-free medium composed of IPL-41 containing 6 g L−1 yeastolate ultrafiltrate, 10 g L−1 glucose, 2 g L−1 lactose, 5 g L−1 glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2 × 107 cells mL−1 in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL−1 and 638 ng RVGP mL−1, respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL−1 and 322 ng HBsAg mL−1 in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3 × 107 cells mL−1 and produced up to 758 ng RVGP mL−1. In general, glucose was consumed by cells, while lactate and ammonia were produced.
KeywordsS2 cells Metabolism RVGP HBsAg Animal protein-free medium
The authors acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, Brazil), Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant no. 02/09482-3, São Paulo, Brazil), Instituto de Pesquisas Tecnológicas do Estado de São Paulo (IPT, São Paulo, Brazil) and Instituto Butantan (São Paulo, Brazil) for the financial support. The authors gratefully acknowledge M Aguiar and R Andrade (IPT, São Paulo, Brazil) for the technical assistance. AM Moraes and CA Pereira are recipients of CNPq fellowships.
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