Document Detail


Microarray and proteomics expression profiling identifies several candidates, including the valosin-containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines.
MedLine Citation:
PMID:  20091739     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A high rate of cell growth (micro) leading to rapid accumulation of viable biomass is a desirable phenotype during scale up operations and the early stages of production cultures. In order to identify genes and proteins that contribute to higher growth rates in Chinese hamster ovary (CHO) cells, a combined approach using microarray and proteomic expression profiling analysis was carried out on two matched pairs of CHO production cell lines that displayed either fast or slow growth rates. Statistical analysis of the microarray and proteomic data separately resulted in the identification of 118 gene transcripts and 58 proteins that were differentially expressed between the fast- and slow-growing cells. Overlap comparison of both datasets identified a priority list of 21 candidates associated with a high growth rate phenotype in CHO. Functional analysis (by siRNA) of five of these candidates identified the valosin-containing protein (VCP) as having a substantial impact on CHO cell growth and viability. Knockdown of HSPB1 and ENO1 also had an effect on cell growth (negative and positive, respectively). Further functional validation in CHO using both gene knockdown (siRNA) and overexpression (cDNA) confirmed that altered VCP expression impacted CHO cell proliferation, indicating that VCP and other genes and proteins identified here may play an important role in the regulation of CHO cell growth during log phase culture and are potential candidates for CHO cell line engineering strategies.
Authors:
Padraig Doolan; Paula Meleady; Niall Barron; Michael Henry; Ross Gallagher; Patrick Gammell; Mark Melville; Martin Sinacore; Kevin McCarthy; Mark Leonard; Timothy Charlebois; Martin Clynes
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  106     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-03-29     Completed Date:  2010-06-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  42-56     Citation Subset:  IM    
Affiliation:
National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland. padraig.doolan@dcu.ie
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphatases / biosynthesis
Animals
CHO Cells
Cell Cycle*
Cell Cycle Proteins / biosynthesis
Cricetinae
Cricetulus
Epithelial Cells / physiology*
Gene Expression*
Gene Expression Profiling*
Gene Silencing
Proteome / analysis*
RNA, Small Interfering / metabolism
Up-Regulation
Chemical
Reg. No./Substance:
0/Cell Cycle Proteins; 0/Proteome; 0/RNA, Small Interfering; EC 3.6.1.-/Adenosine Triphosphatases; EC 3.6.1.-/CDC48 protein

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


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