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

Author:

Saisud Sureewan1ORCID,Posung Manoch2ORCID,Tuntigumthon Suthida2ORCID,Areesirisuk Atsadawut3ORCID,Dhepakson Panadda2ORCID,Teeka Jantima3ORCID

Affiliation:

1. Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Thailand

2. Innovation and Medical Biotechnology Center (iMBC), Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Thailand

3. Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Thailand; Center of Excellence in Nano-Biotechnology and Digital Innovation, Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi, Thailand

Abstract

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.

Publisher

Research Square Platform LLC

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