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Gewählte Publikation:

Hinterkörner, G; Brugger, G; Müller, D; Hesse, F; Kunert, R; Katinger, H; Borth, N.
(2007): Improvement of the energy metabolism of recombinant CHO cells by cell sorting for reduced mitochondrial membrane potential.
J Biotechnol. 2007; 129(4):651-657 FullText FullText_BOKU

One of the major problems in process performance of mammalian cell cultures is the production of lactic acid. Cell specific glucose uptake rates usually correlate to glucose concentration and approximately 80% of the metabolised glucose is converted into lactic acid. As the mitochondrial membrane potential was shown to correlate to cell specific glucose uptake rates, we used Rhodamine 123, a lipophilic cationic dye for cell sorting to improve the energy metabolism of existing production cell lines. Two recombinant CHO cell lines with known differences in lactic acid production rate were used to evaluate Rhodamine 123 staining as a descriptor for glucose uptake rates and to determine whether it is possible to isolate subclones with altered metabolic properties. Such subclones would exhibit an improved process performance, and in addition could be used as models for genomic and metabolic studies. From the cell line with high lactate production, a subclone sorted for reduced mitochondrial membrane potential was found to have a lower specific lactate formation rate compared to the parental cell line in batch cultures. In addition, the glucose consumption rate was also reduced, while both the growth rate and the final cell concentration were increased. A subclone sorted for high mitochondrial membrane potential, on the other hand, had a higher glucose consumption rate, a higher lactate production rate and reduced growth. The potential of using flow cytometric cell sorting methods based on physiological activity for cell line optimisation is discussed. (c) 2007 Elsevier B.V. All rights reserved.
Autor*innen der BOKU Wien:
Borth Nicole
Hesse Friedemann
Hinterkörner Georg
Katinger Hermann
Kunert Renate
Find related publications in this database (using NML MeSH Indexing)
Animals -
CHO Cells -
Cell Separation - methods
Clone Cells - methods
Cricetinae - methods
Cricetulus - methods
Culture Media - methods
Energy Metabolism - methods
Flow Cytometry - methods
Membrane Potentials - physiology
Mitochondrial Membranes - physiology

Find related publications in this database (Keywords)
recombinant Chinese hamster ovary cells
specific lactate production rate
mitochondrial membrane potential
flow cytometry
cell sorting

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