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Effect of CO2 on uninfected Sf-9 cell growth and metabolism

机译：二氧化碳对未感染的Sf-9细胞生长和代谢的影响

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A problem in the mass production of recombinant proteins and biopesticides using insect cell culture is CO2 accumulation. This research investigated the effect of elevated CO2 concentration on insect cell growth and metabolism. Spodoptera frugiperda Sf-9 insect cells were grown at 20% air saturation, 27(degrees)C, and a pH of 6.2. The cells were exposed to a constant CO2 concentration by purging the medium with CO2 and the headspace with air. The population doubling time (PDT) of Sf-9 cells increased with increasing CO2 concentration. Specifically, the PDT for 0-37, 73, 147, 183, and 220 mm Hg CO2 concentrations were 23.2 +/- 6.7, 32.4 +/- 7.2, 38.1 +/- 13.3, 42.9 +/- 5.4, and 69.3 +/- 35.9 h (n=3 or 4, 95% confidence level), respectively. The viability of cells in all experiments was above 90%, i.e., while increased CO2 concentrations inhibited cell growth, it did not affect cell viability. The osmolality for all bioreactor experiments was observed to be 300-360 mOsm/kg, a range that is known to have a negligible effect on insect cell culture. Elevated CO2 concentration did not significantly alter the cell specific glucose consumption rate (2.5-3.2 x 10(-17) mol/cells), but slightly increased the specific lactate production rate from -3.0 x 10(-19) to 10.2 x 10(-19) mol/cells. Oxidative stress did not contribute to CO2 inhibition in uninfected Sf-9 cells as no significant increase in the levels of lipid hydroperoxide and protein carbonyl concentrations was discovered at elevated CO2 concentration. (c) 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:465-469, 2016
机译：使用昆虫细胞培养大规模生产重组蛋白和生物农药的问题是CO2积累。这项研究调查了CO2浓度升高对昆虫细胞生长和代谢的影响。草地贪夜蛾Sf-9昆虫细胞在空气饱和度为20％，27°C和pH值为6.2的条件下生长。通过用二氧化碳吹扫培养基和用空气吹扫顶部空间，使细胞暴露于恒定的二氧化碳浓度。 Sf-9细胞的群体倍增时间（PDT）随着CO2浓度的增加而增加。具体而言，0-37、73、147、183和220 mm Hg CO2浓度的PDT为23.2 +/- 6.7、32.4 +/- 7.2、38.1 +/- 13.3、42.9 +/- 5.4和69.3 + / -35.9小时（n = 3或4，置信度为95％）。在所有实验中，细胞的活力都高于90％，即，虽然增加的CO2浓度抑制了细胞的生长，但它并不影响细胞的活力。观察到所有生物反应器实验的重量克分子渗透浓度为300-360 mOsm / kg，该范围对昆虫细胞培养的影响可忽略不计。升高的CO2浓度不会显着改变细胞比葡萄糖的消耗速率（2.5-3.2 x 10（-17）mol / cells），但比乳酸的产生速率从-3.0 x 10（-19）略微增加到10.2 x 10（ -19）mol / cells。在未感染的Sf-9细胞中，氧化应激对CO2的抑制没有贡献，因为在升高的CO2浓度下，脂质氢过氧化物和蛋白质羰基的浓度没有显着增加。 （c）2016美国化学工程师学会生物技术学会。 Prog。，32：465-469，2016

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