| Effects of conditioned medium factors and passage number on Sf9 cell physiology and productivity. | |
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MedLine Citation:
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PMID: 16599552 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Karin Calles; Ingrid Svensson; Eva Lindskog; Lena Häggström |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biotechnology progress Volume: 22 ISSN: 8756-7938 ISO Abbreviation: Biotechnol. Prog. Publication Date: 2006 Mar-Apr |
Date Detail:
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Created Date: 2006-04-07 Completed Date: 2006-06-09 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8506292 Medline TA: Biotechnol Prog Country: United States |
Other Details:
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Languages: eng Pagination: 394-400 Citation Subset: IM |
Affiliation:
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Karo Bio AB, Huddinge, SE-141 57 Huddinge, Sweden. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Aging / drug effects*, physiology* Cell Line Cell Proliferation / drug effects Culture Media, Conditioned / chemistry*, pharmacology* Molecular Weight Spodoptera / cytology*, drug effects*, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Culture Media, Conditioned |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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