Document Detail

Effects of conditioned medium factors and passage number on Sf9 cell physiology and productivity.
MedLine Citation:
PMID:  16599552     Owner:  NLM     Status:  MEDLINE    
The effects of conditioned medium (CM) and passage number on Spodoptera frugiperda Sf9 cell physiology and productivity have been studied. Low passage (LP) cells at passages 20-45 were compared to high passage (HP) cells at passages >100. Addition of 20% CM or 10 kDa filtrated CM to LP cells promoted growth. LP cells passed a switch in growth kinetics, characterized by a shorter lag phase and a higher growth rate, after 30-40 passages. After this point, CM lost its stimulating effect on proliferation. HP cells displayed a still shorter lag phase and reached the maximum cell density 24-48 earlier than LP cells. HP cells also exhibited higher specific productivity of recombinant protein compared to LP cells, when infected with baculovirus during the initial 48 h of culture. The specific productivity of LP cells was decreased by 30-50% by addition of 20% CM or 10 kDa filtrated CM, whereas addition of CM to cells having passed the switch in growth kinetics had no negative effect on productivity. Cell cycle analysis showed that a large proportion of HP cells, >60%, was transiently arrested in G2/M after inoculation. In LP cultures this proportion was lower, 40-45%, and addition of CM decreased the arrested population further. This correlated to the cell size, the HP cells being the largest: HP cells > LP > LP + 20% CM > LP + 20% 10 kDa filtrated CM. Since the degree of synchronization in G2/M correlated to the productivity, yeastolate limitation was used to achieve 85% G2/M synchronized cells. In this culture the specific productivity was maintained during a prolonged production phase and a 69% higher volumetric yield was obtained. The results suggest that a decreasing degree of synchronization during the course of culture partly explains the cell-density-dependent drop in productivity in Sf9 cells.
Karin Calles; Ingrid Svensson; Eva Lindskog; Lena Häggström
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 Mar-Apr
Date Detail:
Created Date:  2006-04-07     Completed Date:  2006-06-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  394-400     Citation Subset:  IM    
Karo Bio AB, Huddinge, SE-141 57 Huddinge, Sweden.
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MeSH Terms
Cell Aging / drug effects*,  physiology*
Cell Line
Cell Proliferation / drug effects
Culture Media, Conditioned / chemistry*,  pharmacology*
Molecular Weight
Spodoptera / cytology*,  drug effects*,  metabolism
Reg. No./Substance:
0/Culture Media, Conditioned

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

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