Development of an Animal-Derived Component-Free Medium for Spodoptera frugiperda (Sf9) Cells Using Response Surface Methodology

Abstract Spodoptera frugiperda (Sf9) insect cells have been widely used for recombinant protein production. Currently, serum-free media for Sf9 cells are commercially available, but they mostly still contain animal-derived components. This work aimed to develop an animal-derived component free medium for Sf9 cells. Chemically defined lipid concentrate (CDLC), yeast extract (YE), and soytone (ST) were studied for serum substitutes using a central composite design (CCD). OSF9-ADCFM contained optimum concentrations of CDLC, YE and ST at 0.5% (v/v), 11.0 g/L, and 3.0 g/L, respectively. The mean viable cell concentration of 1.71 ± 0.14 x 105 cells/mL was obtained from 5 passages (P1-P5) in 25 cm2 tissue culture flask. The use of both peptones after 10-kDa ultrafiltration had a significant effect on Sf9 cells growth. Maximum cell concentrations and the specific growth rates of cell adaptation in suspension culture were 1.79–2.17 x 106 cells/mL and 0.025–0.027 h− 1, respectively. The results indicated that OSF9-ADCFM could be used for Sf9 culture in both static and suspension culture. Further study would be needed to verify the effect on protein production of this in-house animal-derived component-free medium.