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

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논문

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

학술지

Biotechnology progress

저자명

Vajrala, Sucheta G.; Murhammer, David W.

초록

<P>A problem in the mass production of recombinant proteins and biopesticides using insect cell culture is CO<SUB>2</SUB> accumulation. This research investigated the effect of elevated CO<SUB>2</SUB> concentration on insect cell growth and metabolism. Spodoptera frugiperda Sf&#8208;9 insect cells were grown at 20% air saturation, 27<SUP>&deg;</SUP>C, and a pH of 6.2. The cells were exposed to a constant CO<SUB>2</SUB> concentration by purging the medium with CO<SUB>2</SUB> and the headspace with air. The population doubling time (PDT) of Sf&#8208;9 cells increased with increasing CO<SUB>2</SUB> concentration. Specifically, the PDT for 0&#8208;37, 73, 147, 183, and 220 mm Hg CO<SUB>2</SUB> concentrations were 23.2 &plusmn; 6.7, 32.4 &plusmn; 7.2, 38.1 &plusmn; 13.3, 42.9 &plusmn; 5.4, and 69.3 &plusmn; 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 CO<SUB>2</SUB> concentrations inhibited cell growth, it did not affect cell viability. The osmolality for all bioreactor experiments was observed to be 300&ndash;360 mOsm/kg, a range that is known to have a negligible effect on insect cell culture. Elevated CO<SUB>2</SUB> concentration did not significantly alter the cell specific glucose consumption rate (2.5&ndash;3.2 &times; 10<SUP>&minus;17</SUP> mol/cell s), but slightly increased the specific lactate production rate from &minus;3.0 &times; 10<SUP>&minus;19</SUP> to 10.2 &times; 10<SUP>&minus;19</SUP> mol/cell s. Oxidative stress did not contribute to CO<SUB>2</SUB> inhibition in uninfected Sf&#8208;9 cells as no significant increase in the levels of lipid hydroperoxide and protein carbonyl concentrations was discovered at elevated CO<SUB>2</SUB> concentration. &copy; 2016 American Institute of Chemical Engineers <I>Biotechnol. Prog.</I>, 32:465&ndash;469, 2016</P>

발행연도

2016

ISSN

8756-7938

ISSN

1520-6033

32

2

페이지

pp.465-469

주제어

Spodoptera frugiperda Sf&#x2010; 9 cells; carbon dioxide inhibition; metabolism; cell growth;

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논문; 2016-12-31

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