Effect of conditioned medium factors on productivity and cell physiology in Trichoplusia ni insect cell cultures.
2006 (English)In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 22, no 3, 653-659 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2006. Vol. 22, no 3, 653-659 p.
recombinant protein-production, baculovirus expression system, fed-batch culture, medium replacement, sf-9 cells, growth, line, suspension, infection, oxygen
IdentifiersURN: urn:nbn:se:kth:diva-5221DOI: 10.1021/bp050252+ISI: 000237973500007ScopusID: 2-s2.0-33745027387OAI: oai:DiVA.org:kth-5221DiVA: diva2:8042
Tidigare titel: Conditioned medium factors regulate productivity and cell physiology in Trichoplusia ni insect cell cultures.
Uppdaterad från submitted till published(20101125)
QC 201011252005-05-312005-05-312011-02-15Bibliographically approved