Cytotechnology

, Volume 22, Issue 1–3, pp 225–237 | Cite as

The effect of dissolved oxygen tension and the utility of oxygen uptake rate in insect cell culture

  • Laura A. Palomares
  • Octavio T. Ramirez
Special Issue

Abstract

Dissolved oxygen tension and oxygen uptake rate are critical parameters in animal cell culture. However, only scarce information of such variables is available for insect cell culture. In this work, the effect of dissolved oxygen tension (DOT) and the utility of on-line oxygen uptake rate (OUR) measurements in monitoring Spodoptera frugiperda (Sf9) cultures were determined. Sf9 cells were grown at constant dissolved oxygen tensions in the range of 0 to 30%. Sf9 metabolism was affected only at DOT below 10%, as no significant differences on specific growth rate, cell concentration, amino acid consumption/production nor carbohydrates consumption rates were found at DOT between 10 and 30%. The specific growth rate and specific oxygen uptake rate followed typical Monod kinetics with respect to DOT. The calculated μmax and \(q_{O_2 }\) max were 0.033 h-1 and 3.82×10-10 mole cell-1h-1, respectively, and the corresponding saturation constants were 1.91 and 1.57%, respectively. In all aerated cultures, lactate was consumed only after glucose and fructose had been exhausted. The yield of lactate increased with decreasing DOT. It is proposed, that an ‘apparent’ DOT in non-instrumented cultures can be inferred from the lactate yield of bioreactors as a function of DOT. Such a concept, can be a useful and important tool for determining the average dissolved oxygen tension in non-instrumented cultures. It was shown that the dynamic behavior of OUR can be correlated with monosaccharide (fructose and glucose) depletion and viable cell concentration. Accordingly, OUR can have two important applications in insect cell culture: for on-line estimation of viable cells, and as a possible feed-back control variable in automatic strategies of nutrient addition.

Key words

Insect cells dissolved oxygen tension oxygen uptake rate metabolism aminoacids carbohydrates 

Abbreviations

DOT

Dissolved oxygen tension

OUR

Oxygen uptake rate

\(q_{O_2 }\)

specific oxygen uptake rate

μ

specific growth rate

Xv

viable cell concentration

CL, C*, and \(C_{H_2 O}\)

oxygen concentrations in liquid phase, in equilibrium with gas phase, and medium molar concentration, respectively

H

Henry's constant

KLa

volumetric oxygen transfer coefficient

PT

total pressure

\(p_{O_2 }\)

oxygen partial pressure

\(y_{O_2 }\)

oxygen molar fraction

i

discrete element

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Laura A. Palomares
    • 1
  • Octavio T. Ramirez
    • 1
  1. 1.Departamento de Bioingeniería, Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMéxico

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