Towards a metabolic and isotopic steady state in CHO batch cultures for reliable isotope-based metabolic profiling

被引：37

作者：

Biochemical Engineering Institute, Saarland University, Bldg. 2, 66123 Saarbrücken, Germany ^{[1
]}

不详 ^{[2
]}

不详 ^{[3
]}

不详 ^{[4
]}

机构：

[1] Biochemical Engineering Institute, Saarland University, 66123 Saarbrücken

[2] James Graham Brown Cancer Center, Department of Surgery, University of Louisville, Louisville, KY

[3] Laboratory of Chemical Biotechnology, Faculty of Biochemical and Chemical Engineering, TU Dortmund, 44221 Dortmund

[4] Genomatica, Inc., San Diego, CA 92121

来源：

Biotechnol. J. | 2009年 / 2卷 / 247-263期

关键词：

Balanced growth; CHO cells; Isotopic labeling; Media optimization; Metabolic profiling;

D O I：

10.1002/biot.200800143

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Attaining metabolic and isotopic balanced growth is one critical condition for physiological studies using isotope-labeled tracers, but is very difficult to obtain in batch culture due to the extensive metabolite exchange with the surrounding medium and related physiological changes. In the present study, we investigated metabolic and isotopic behavior of CHO cells in differently designed media. We observed that the assumption of balanced cell growth cannot be justified in batch culture of CHO cells directly using conventional, commercially available media. By systematically redesigning media composition and characterizing metabolic steady state based on mass balances and measurement of labeling dynamics, we achieved balanced cell growth for the main cellular substrates in CHO cells. This was done in a step-by-step analysis of growth and primary metabolism of CHO cells with the use of [U-13C]glucose feeding and adjusting concentrations of amino acids in the growth medium. The optimized media obtained at the end of the study provide balanced growth and isotopic steady state or at least asymptotic steady state. As a result, we established a platform to conduct isotope-based physiological studies of mammalian systems more reliably and therefore well suited for later use in metabolic profiling of mammalian systems such as 13C-labeled metabolic flux analysis. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

引用

页码：247 / 263

页数：16

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