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Retrovirus producer cell line metabolism: implications on viral productivity
Retrovirus producer cell line metabolism: implications on viral productivity
- Source :
- Applied Microbiology and Biotechnology. 72:1125-1135
- Publication Year :
- 2006
- Publisher :
- Springer Science and Business Media LLC, 2006.
-
Abstract
- The production of retroviral vectors by human cell lines is still hampered by low titers making it relatively difficult to produce very large quantities of this vector of high interest for clinical gene therapy applications. Thus, to improve vector production, we studied the influence of different sugars alone or combinations of sugars on cell growth, vector titers, and metabolism of the producer cell. The use of fructose at 140 mM or a mixed medium (with glucose at 25 mM and fructose at 140 mM) improved the virus titer three- to fourfold, respectively, and the producer cell productivity by fivefold. The increase in the cell productivity was due to a 1.5-fold increase in the vector stability, the remaining increase being due to higher cell specific productivity. The increase in the productivity was associated with lower glucose oxidation and an increase in the lactate and alanine yield. In the mixed medium, an increase in fatty acids derived from the glucose was observed in parallel with a reduction of glutamate and glutamine synthesis via the tricarboxylic acid (TCA) cycle acetyl-CoA and alpha-ketoglutarate, respectively. Although the higher productivities were associated with severe changes in the glycolysis, TCA cycle, and glutaminolysis, the cell energetic status monitored by phosphocreatine and adenosine triphosphate levels was not significantly affected. The synthesis of fatty acids and phospholipids were enhanced in the fructose or mixed media and are possibly key parameters in retroviral vector production.
- Subjects :
- Magnetic Resonance Spectroscopy
Virus Cultivation
Phosphocreatine
Glutamine
Genetic Vectors
Glutamic Acid
Cell Count
Fructose
Carbohydrate metabolism
Biology
Applied Microbiology and Biotechnology
chemistry.chemical_compound
Adenosine Triphosphate
Cell Line, Tumor
Humans
Glycolysis
Lactic Acid
Alanine
Glutaminolysis
Cell growth
Fatty Acids
General Medicine
Metabolism
Culture Media
Citric acid cycle
Glucose
Retroviridae
chemistry
Biochemistry
Oxidation-Reduction
Biotechnology
Subjects
Details
- ISSN :
- 14320614 and 01757598
- Volume :
- 72
- Database :
- OpenAIRE
- Journal :
- Applied Microbiology and Biotechnology
- Accession number :
- edsair.doi.dedup.....4cf1d288ad5b0cbd113c656ae735055d
- Full Text :
- https://doi.org/10.1007/s00253-006-0401-y