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Generation of stable, high-producing cho cell lines by lentiviral vector-mediated gene transfer in serum-free suspension culture.
MedLine Citation:
PMID:  20967750     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Lentivirus-derived vectors (LVs) were studied for the generation of stable recombinant Chinese hamster ovary (CHO) cell lines. Stable pools and clones expressing the enhanced green fluorescent protein (eGFP) were selected via fluorescence-activated cell sorting (FACS). For comparison, cell pools and cell lines were also generated by transfection, using the LV transfer plasmid alone. The level and stability of eGFP expression was greater in LV-transduced cell lines and pools than in those established by transfection. CHO cells were also infected at two different multiplicities of infection with an LV co-expressing eGFP and a tumor necrosis factor receptor:Fc fusion protein (TNFR:Fc). At 2-day post-infection, clonal cell lines with high eGFP-specific fluorescence were recovered by FACS. These clones co-expressed TNFR:Fc with yields of 50-250 mg/L in 4-day cultures. The recovered cell lines maintained stable expression over 3 months in serum-free suspension culture without selection. In conclusion, LV-mediated gene transfer provided an efficient alternative to plasmid transfection for the generation of stable and high-producing recombinant cell lines. Biotechnol. Bioeng. 2011; 108:600-610. © 2010 Wiley Periodicals, Inc.
Authors:
Agata Oberbek; Mattia Matasci; David L Hacker; Florian M Wurm
Publication Detail:
Type:  Journal Article     Date:  2010-11-23
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  108     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-01-19     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  600-10     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Wiley Periodicals, Inc.
Affiliation:
Ecole Polytechnique Fédérale de Lausanne (EPFL), Laboratory of Cellular Biotechnology, CH-1015 Lausanne, Switzerland; telephone: +41-21-693-61-41; fax: +41-21-693-61-40.
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