Document Detail


Metabolism of PER.C6 cells cultivated under fed-batch conditions at low glucose and glutamine levels.
MedLine Citation:
PMID:  16523524     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Luis Maranga; Charles F Goochee
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  94     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2006 May 
Date Detail:
Created Date:  2006-04-19     Completed Date:  2006-06-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  139-50     Citation Subset:  IM    
Affiliation:
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
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MeSH Terms
Descriptor/Qualifier:
Adenoviridae / 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:
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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