Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media

Abstract

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.