Document Detail


Efficient transfection of primary cells relevant for cardiovascular research by nucleofection.
MedLine Citation:
PMID:  17085816     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cell types that are important for cardiovascular research, e.g., cardiomyocytes, endothelial cells, or adult stem cells, are often hard to isolate, culture, and transfect. Low-transfection efficiencies are a major limitation because, in many cases, results achieved with surrogate model cell lines, if any at all are available for the primary cell type of interest, do not reflect the situation in the primary cell. We have demonstrated that unprecedented transfection results are achieved with primary cells when novel electroporation conditions are combined with a treatment of the cells in specific solutions that help stabilize the cells in the electrical field. This led to the development of the new proprietary transfection technology nucleofection. Nucleofection has proved to be successfully applicable to a variety of primary cells and other hard-to-transfect cell lines, and, thus, opens unique perspectives for novel experimental setups as therapeutic strategies. Herein we present protocols for the efficient nucleofection of human umbilical vein endothelial cells, human coronary artery endothelial cells, smooth muscle cells (e.g., pig vascular smooth muscle cells), neonatal rat cardiomyocytes, and human mesenchymal stem cells and depict some results obtained with such transfected cells.
Authors:
Corinna Thiel; Michael Nix
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular medicine     Volume:  129     ISSN:  1543-1894     ISO Abbreviation:  Methods Mol. Med.     Publication Date:  2006  
Date Detail:
Created Date:  2006-11-06     Completed Date:  2006-12-20     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101123138     Medline TA:  Methods Mol Med     Country:  United States    
Other Details:
Languages:  eng     Pagination:  255-66     Citation Subset:  IM    
Affiliation:
Amaxa Biosystems, Cologne, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques
Coronary Vessels / cytology
Electroporation / methods
Endothelial Cells*
Flow Cytometry
Fluorescent Antibody Technique
Humans / genetics
Mesenchymal Stem Cells*
Microscopy, Fluorescence
Myocytes, Cardiac*
Myocytes, Smooth Muscle*
Rats / genetics
Transfection / methods*
Umbilical Veins / cytology

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


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