Document Detail


Effect of conditioned medium factors on productivity and cell physiology in Trichoplusia ni insect cell cultures.
MedLine Citation:
PMID:  16739946     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The influence of conditioned medium (CM) on cell physiology and recombinant protein production in Trichoplusia ni insect cells (T. ni, BTI-Tn-5B1-4) has been investigated. Cell cycle analysis showed that a high proportion of the cell population (80-90%) was in G1 during the whole culture, indicating that the S and G2/M phases are short relative to the G1 phase. Directly after inoculation, a rapid decrease of the S-phase population occurred, which could be observed as a lag-phase. The following increase in the number of cells in S occurred after 7 h of culture for cells in fresh medium, whereas for cells with the addition of CM it occurred at an earlier time point (5 h) and these cells had therefore a shorter lag-phase. The initial changes in the S-phase population were also affected by the inoculum cell density, as higher seeding cell densities resulted in a more rapid increase in the S-phase population after inoculation. These changes in cell cycle distribution were reflected in the cell size, and the CM-cells were smaller than the cells in fresh medium. Recombinant protein production in T. ni cells was improved by the addition of CM. The specific productivity was increased by 30% compared to cells in fresh medium. This beneficial effect was seen between 20 and 72 h of culture. In contrast, the highest specific productivity was obtained already at 7 h for the cells in fresh medium and then decreased rapidly. The total product concentration was around 30% higher in the culture with CM compared to the culture in fresh medium, and the maximum product concentration was obtained on day 2 compared to day 3 for the cells in fresh medium. Our results indicate that the positive effect on productivity by CM is related to its growth-promoting effect, suggesting that the proliferation potential of the culture determines the productivity.
Authors:
Karin Calles; Ulrika Eriksson; Lena Häggström
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology progress     Volume:  22     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2006 May-Jun
Date Detail:
Created Date:  2006-06-02     Completed Date:  2006-08-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  653-9     Citation Subset:  IM    
Affiliation:
Karo Bio AB, Huddinge, SE-141 57 Huddinge, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques / methods
Cell Cycle / drug effects
Cell Line
Cell Proliferation / drug effects
Cells, Cultured
Culture Media, Conditioned / pharmacology*
Insect Proteins / biosynthesis*,  drug effects
Moths / cytology,  metabolism*
Recombinant Proteins / biosynthesis,  drug effects
Time Factors
Chemical
Reg. No./Substance:
0/Culture Media, Conditioned; 0/Insect Proteins; 0/Recombinant Proteins

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


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