Document Detail


Optimization of environment for high density Vero cell culture: effect of dissolved oxygen and nutrient supply on cell growth and changes in metabolites.
MedLine Citation:
PMID:  3733899     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This study was initiated for optimization of the environment of a technologically useful mammalian cell line for high density production. Cultures of Vero cells on microcarriers were perfused with 100%, 50%, 25% and 12.5% modified L15 media (galactose was replaced with 10 mM-fructose, with 4 mM-glutamine and 5% foetal bovine serum) in phosphate-buffered saline at either 4 or 8 vol. day-1. Cell growth, pH, dissolved oxygen, and changes in the metabolites, lactate to pyruvate and lactate to ammonia indices, demonstrated that under the conditions used in the present study, perfusion of cultures with 50% L15 medium in PBS at 8 vol. day-1 provided the optimum microenvironment for Vero cell growth. The highest cell density in the perfused cultures was 3 X 10(7) cells ml-1, which at these conditions was ten times higher than the maximum cell density (3 X 10(6) cells ml-1) obtained in a batch culture. Nutrient supply and conditioning factors were the most probable growth-limiting factors in cultures that were perfused with 12.5% and 25% L15 media, while multilayering, limitation of available oxygen, and accumulation of metabolic end products in the cellular microenvironment were the most probable causes of a density-dependent inhibition of cell growth observed under the optimized and overfed (supply of 100% L15 medium at the rate of 8 vol. day-1) culture conditions. Under the optimized environmental condition, the major source of energy was probably glutamine during the first week. However, significant utilization of fructose became evident at higher cell densities during the second week, when lactate production dramatically declined and reached an almost undetectable level, while respiration progressively assumed the predominant role in energy production. It is postulated that 'available' oxygen in the multicell-layered microenvironment of the optimized cultures was higher than in the overfed culture due to the greater utilization rate of oxygen for oxidation of excess nutrients in the overfed culture.
Authors:
A T Nahapetian; J N Thomas; W G Thilly
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of cell science     Volume:  81     ISSN:  0021-9533     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  1986 Mar 
Date Detail:
Created Date:  1986-09-17     Completed Date:  1986-09-17     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  65-103     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Ammonia / metabolism
Animals
Cell Count
Cell Line
Cercopithecus aethiops
Culture Media*
Glucose / metabolism
Hydrogen-Ion Concentration
Kidney / cytology
Lactates / metabolism
Mitosis*
Oxygen / metabolism
Pyruvates / metabolism
Time Factors
Chemical
Reg. No./Substance:
0/Culture Media; 0/Lactates; 0/Pyruvates; 50-99-7/Glucose; 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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