Division of Biology, Faculty of Science and Technology, Rajamangala University of Technology, Thanyaburi, Thailand.
Innovation and Medical Biotechnology Center (iMBC), Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
Biotechnol Lett. 2023 Jul;45(7):761-777. doi: 10.1007/s10529-023-03389-5. Epub 2023 May 15.
To develop an animal-derived component-free medium for Spodoptera frugiperda (Sf9) growth and green fluorescent protein (GFP) expression.
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 × 10 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.
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.
开发一种不含动物源性成分的培养基用于 S. frugiperda（Sf9）生长和绿色荧光蛋白（GFP）表达。
OSF9-ADCFM 含有最佳浓度的 CDLC、YE 和 ST，分别为 0.5%（v/v）、11.0 g/L 和 3.0 g/L。在 5 个传代（P1-P5）中获得了 1.71±0.14×10^6 个/mL 的平均活细胞浓度。10 kDa 超滤后使用两种蛋白胨对 Sf9 细胞生长有显著影响。Grace 昆虫培养基添加 10% FBS 产生的未感染细胞数比 SF-900II 和 OSF9-ADCFM 分别高 4.29 倍和 5.38 倍。SF-900II 中未感染细胞和 GFP 荧光细胞的平均细胞数分别比 OSF9-ADCFM 高 1.25 倍和 7 倍。
自制的 OSF9-ADCFM 可以支持 Sf9 的生长和 GFP 表达，但不如商业的 SF-900II。然而，与商业 SF-900II 相比，它可以将生产成本降低至少 50%。下一步将继续开发自制的 OSF9-ADCFM，以提高细胞数量和蛋白表达。
