Document Detail


Targeting early apoptotic genes in batch and fed-batch CHO cell cultures.
MedLine Citation:
PMID:  16894638     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Based on the transcriptional profiling of CHO cell culture using microarray, four key early apoptosis signaling genes, Fadd, Faim, Alg-2, and Requiem, were identified and CHO GT (Gene Targeted) cell lines were developed by targeting these four genes. Two were CHO GT(O) cell lines overexpressing anti-apoptotic genes, Faim and Fadd DN and two were CHO GT(KD) cell lines involving knockdown of Alg-2 and Requiem which are pro-apoptotic genes using small interfering RNA (siRNA) technology. Comparisons of these CHO GT cell lines with the parental cell line in batch culture (BC) and fed-batch culture (FBC) were performed. Compared to parental cells, the CHO GT cell lines showed apoptosis resistance as they significantly delayed and/or suppressed initiator caspase-8 and -9 and executioner caspase-3 activities during culture. FBC of CHO GT cell lines reached significantly higher maximum viable cell densities (up to 9 x 10(6) cells/mL) compared with the parental cell line (5 x 10(6) cells/mL). The recombinant interferon gamma (IFN-gamma) yields were increased by up to 2.5-fold. Furthermore, it was observed that the IFN-gamma was more highly sialylated.
Authors:
Danny Chee Furng Wong; Kathy Tin Kam Wong; Peter Morin Nissom; Chew Kiat Heng; Miranda Gek Sim Yap
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  95     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2006 Oct 
Date Detail:
Created Date:  2006-09-04     Completed Date:  2006-11-09     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  350-61     Citation Subset:  IM    
Affiliation:
Bioprocessing Technology Institute, Biomedical Sciences Institutes, Agency for Science, Technology and Research, A*STAR, 20 Biopolis Way, #06-01, Centros, Singapore 138668.
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MeSH Terms
Descriptor/Qualifier:
Adaptor Proteins, Signal Transducing / genetics
Animals
Apoptosis / genetics*,  physiology
Apoptosis Regulatory Proteins / genetics*,  metabolism
Bioreactors*
CHO Cells
Calcium-Binding Proteins / genetics
Carrier Proteins / genetics
Cell Cycle Proteins / genetics
Cricetinae
Cricetulus
DNA-Binding Proteins / genetics
Down-Regulation / genetics
Endosomal Sorting Complexes Required for Transport
Fas-Associated Death Domain Protein
Gene Expression Profiling / methods
Interferon-gamma / metabolism
Oligonucleotide Array Sequence Analysis / methods
Recombinant Proteins / metabolism
Chemical
Reg. No./Substance:
0/Adaptor Proteins, Signal Transducing; 0/Apoptosis Regulatory Proteins; 0/Calcium-Binding Proteins; 0/Carrier Proteins; 0/Cell Cycle Proteins; 0/DNA-Binding Proteins; 0/DPF2 protein, human; 0/Endosomal Sorting Complexes Required for Transport; 0/FADD protein, human; 0/FAIM protein, human; 0/Fas-Associated Death Domain Protein; 0/PDCD6IP protein, human; 0/Recombinant Proteins; 82115-62-6/Interferon-gamma

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


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