| Metabolism of PER.C6 cells cultivated under fed-batch conditions at low glucose and glutamine levels. | |
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MedLine Citation:
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PMID: 16523524 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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This is the first study to examine PER.C6 cell glucose/energy and glutamine metabolism with fed-batch cultures at controlled low glutamine, low glucose, and simultaneous low glucose and low glutamine levels. PER.C6(TM) cell metabolism was investigated in serum-free suspension bioreactors at two-liter scale. Control of glucose and/or glutamine concentrations had a significant effect on cellular metabolism leading to an increased efficiency of nutrient utilization, altered byproduct synthesis, while having no effect on cell growth rate. Cultivating cells at a controlled glutamine concentration of 0.25 mM reduced q(Gln) and q(NH(4)(+)) by approximately 30%, q(Ala) 85%, and q(NEAA) 50%. The fed-batch control of glutamine also reduced the overall accumulation of ammonium ion by approximately 50% by minimizing the spontaneous chemical degradation of glutamine. No major impact upon glucose/energy metabolism was observed. Cultivating cells at a glucose concentration of 0.5 mM reduced q(Glc) about 50% and eliminated lactate accumulation. Cells exhibited a fully oxidative metabolism with Y(O(2)/Glc) of approximately 6 mol/mol. However, despite no increase in q(Gln), an increased ammonium ion accumulation and Y(NH(4)(+)/Gln) were also observed. Effective control of lactate and ammonium ion accumulation by PER.C6 cells was achieved using fed-batch with simultaneously controlled glucose and glutamine. A fully oxidative glucose metabolism and a complete elimination of lactate production were obtained. The q(Gln) value was again reduced and, despite an increased q(NH(4)(+)) compared with batch culture, ammonium ion levels were typically lower than corresponding ones in batch cultures, and the accumulation of non-essential amino acids (NEAA) was reduced about 50%. In conclusion, this study shows that PER.C6 cell metabolism can be confined to a state with improved efficiencies of nutrient utilization by cultivating cells in fed-batch at millimolar controlled levels of glucose and glutamine. In addition, PER.C6 cells fall into a minority category of mammalian cell lines for which glutamine plays a minor role in energy metabolism. |
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Authors:
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Luis Maranga; Charles F Goochee |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Biotechnology and bioengineering Volume: 94 ISSN: 0006-3592 ISO Abbreviation: Biotechnol. Bioeng. Publication Date: 2006 May |
Date Detail:
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Created Date: 2006-04-19 Completed Date: 2006-06-15 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7502021 Medline TA: Biotechnol Bioeng Country: United States |
Other Details:
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Languages: eng Pagination: 139-50 Citation Subset: IM |
Affiliation:
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Fermentation and Cell Culture, Bioprocess R&D, Merck Research Laboratories, Merck & Co., Inc., 770 Sumneytown Pike, WP17-201 P.O. Box 4, West Point, Pennsylvania 19486, USA. luis_maranga@merck.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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genetics Amino Acids / metabolism Ammonia / metabolism Bioreactors* Cell Count Cell Culture Techniques Cell Line, Transformed Cell Survival Cell Transformation, Viral Culture Media / chemistry Culture Media, Serum-Free Energy Metabolism Glucose / metabolism* Glutamine / metabolism* Humans Lactic Acid / metabolism Oxygen / metabolism Retina / cytology, embryology, growth & development*, metabolism*, virology Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Amino Acids; 0/Culture Media; 0/Culture Media, Serum-Free; 50-21-5/Lactic Acid; 50-99-7/Glucose; 56-85-9/Glutamine; 7664-41-7/Ammonia; 7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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