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Development of an animal-derived component-free medium for Spodoptera frugiperda (Sf9) cells using response surface
Sureewan Saisud1, Manoch Posung2, Suthida Tuntigumthon2
1Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology, Thanyaburi, Thailand.
Objectives:
To develop an animal-derived component-free medium for Spodoptera frugiperda (Sf9) growth and green fluorescent protein (GFP) expression.
Results:
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. A mean viable cell concentration of 1.71 ± 0.14 × 105 cells/mL was obtained from 5 passages (P1-P5). The use of both peptones after 10 kDa ultrafiltration had a significant effect on Sf9 cell growth. Grace's insect medium with 10% FBS gave higher un-infected cell number than SF-900II and OSF9-ADCFM for 4.29 and 5.38 times, respectively. The average cell number of un-infected cells and GFP-fluorescent cells of SF-900II were higher than OSF9-ADCFM 1.25 and 7 times, respectively.
Conclusion:
In-house OSF9-ADCFM could support growth and GFP expression in Sf9 less than commercial SF-900II. However, it could lower the production cost at least 50% comparing to commercial SF-900II. The development of in- house OSF9-ADCFM would be continued to increase both cell numbers and protein expression in the next step.
