Document Detail


High efficiency transfection of glioma cell lines and primary cells for overexpression and RNAi experiments.
MedLine Citation:
PMID:  16621008     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In order to investigate the impact of signalling proteins on the phenotype and malignant behavior of glioblastoma cells, we optimized the transfection procedure of human glioblastoma cell lines U251, U373, GaMG and of primary cells obtained from a patient's tumor using nucleofection technology in conjunction with plasmid pmaxGFP. We describe the optimization procedure, show that a high percentage of the cells can be transfected and that nucleofection does not cause phenotypic alterations of the cells. Therefore, we conclude that nucleofection is a highly efficient tool to deliver plasmids for transient protein overexpression and siRNA for specific protein knock-down to different glioblastoma cell lines or primary cells.
Authors:
Carsten Hagemann; Christoph Meyer; Jelena Stojic; Sven Eicker; Stefanie Gerngras; Siglinde Kühnel; Klaus Roosen; Giles Hamilton Vince
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-04-18
Journal Detail:
Title:  Journal of neuroscience methods     Volume:  156     ISSN:  0165-0270     ISO Abbreviation:  J. Neurosci. Methods     Publication Date:  2006 Sep 
Date Detail:
Created Date:  2006-08-24     Completed Date:  2006-11-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7905558     Medline TA:  J Neurosci Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  194-202     Citation Subset:  IM    
Affiliation:
Department of Neurosurgery, Tumorbiology Laboratory, University of Würzburg, Josef-Schneider-Str. 11, D-97080 Würzburg, Germany. hagemann_c@klinik-uni-wuerzburg.de
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MeSH Terms
Descriptor/Qualifier:
Blotting, Western
Brain Neoplasms / genetics*,  metabolism*
Cell Line, Tumor
Cell Movement
Cell Nucleus / genetics
Cell Proliferation
Cytological Techniques
Electroporation
Glial Fibrillary Acidic Protein / biosynthesis,  genetics
Glioma / genetics*,  metabolism*
Green Fluorescent Proteins
Humans
Immunohistochemistry
Neurons / metabolism*
Phenotype
Plasmids / genetics
RNA, Small Interfering / biosynthesis*
Reverse Transcriptase Polymerase Chain Reaction
Transfection / methods*
Chemical
Reg. No./Substance:
0/Glial Fibrillary Acidic Protein; 0/RNA, Small Interfering; 147336-22-9/Green Fluorescent Proteins

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


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