Document Detail


Optimal NS0 cell growth in a hollow fiber bioreactor requires increased serum concentration or a cholesterol supplement on the cell side of the fiber.
MedLine Citation:
PMID:  14656153     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An NSO/GS cell line secreting a humanized antibody was routinely propagated in a T-flask using 2% serum. For scale-up of antibody production, this cell line was inoculated into a hollow fiber system using the same serum concentration. The metabolic activity increased for a few days in the hollow fiber system, but invariably the activity dropped dramatically as the cells died by day 7. A hollow fiber micro-bioreactor was used as a screening tool to examine possible reasons for cell death in the large-scale system. As seen in the hollow fiber system, cells died when 2% serum was used either on the cell side only or on both sides of the fiber in the micro-bioreactor. In contrast, the use of 20% serum on the cell side of the fiber and basal medium on the non-cell side resulted in good cell expansion at high viability. Regardless of the cell side serum concentration, no further growth enhancements were seen when up to 20% serum was placed on the non-cell side of the fiber. These results suggest that a serum component that does not readily cross the fiber is limiting cell growth in the hollow fiber bioreactors. The addition of a cholesterol-rich lipid supplement resulted in better cell growth in the micro-bioreactor, while the addition of other non-cholesterol lipid supplements resulted in no growth enhancement. The growth-enhancing properties of the cholesterol supplement were more pronounced at lower serum concentrations, suggesting that poor growth at low serum concentration was due to suboptimal cholesterol levels. When the cell side serum concentration was increased to 20% in the hollow fiber system, cells grew and filled the bioreactor, allowing a 39-day production run. These results demonstrate that this NSO cell line requires an increased cell side serum concentration for optimal growth and that this requirement is likely due to the inherent cholesterol dependency of this cell line.
Authors:
Michael J Gramer; Jodi Maas
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article    
Journal Detail:
Title:  Biotechnology progress     Volume:  19     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2003 Nov-Dec
Date Detail:
Created Date:  2003-12-05     Completed Date:  2004-08-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1762-6     Citation Subset:  IM    
Affiliation:
BioVest International, Inc., 8500 Evergreen Boulevard, Minneapolis, Minnesota 55433, USA. mgramer@biovest.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies / metabolism*
Bioreactors*
Cell Culture Techniques / instrumentation,  methods*
Cell Division
Cell Line, Tumor
Cell Survival
Cholesterol / metabolism*
Culture Media / metabolism
Mice
Multiple Myeloma / metabolism*,  pathology*
Pilot Projects
Serum / metabolism*
Ultrafiltration / instrumentation,  methods*
Chemical
Reg. No./Substance:
0/Antibodies; 0/Culture Media; 57-88-5/Cholesterol

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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