Development of an animal-derived component-free medium for Spodoptera frugiperda (Sf9) cells using response surface methodology

被引:3
|
作者
Saisud, Sureewan [1 ]
Posung, Manoch [2 ]
Tuntigumthon, Suthida [2 ]
Areesirisuk, Atsadawut [1 ,3 ]
Dhepakson, Panadda [2 ]
Teeka, Jantima [1 ,3 ]
机构
[1] Rajamangala Univ Technol, Fac Sci & Technol, Div Biol, Thanyaburi, Thailand
[2] Minist Publ Hlth, Dept Med Sci, Med Life Sci Inst, Innovat & Med Biotechnol Ctr iMBC, Nonthaburi, Thailand
[3] Rajamangala Univ Technol, Fac Sci & Technol, Ctr Excellence Nanobiotechnol & Digital Innovat, Thanyaburi, Thailand
关键词
Animal-derived component-free medium; Central composite design; Chemically defined lipid concentrate; Green fluorescent protein; Spodoptera frugiperda (Sf9); Soytone; Yeast extract; SERUM-FREE MEDIA; FETAL BOVINE SERUM; INSECT CELLS; YEASTOLATE FRACTIONS; CULTURE-MEDIA; GROWTH; PEPTONE; EXPRESSION; PROLIFERATION; HYDROLYSATE;
D O I
10.1007/s10529-023-03389-5
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
ObjectivesTo develop an animal-derived component-free medium for Spodoptera frugiperda (Sf9) growth and green fluorescent protein (GFP) expression.ResultsOSF9-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 x 10(5) 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.ConclusionIn-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.
引用
收藏
页码:761 / 777
页数:17
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