Linked Life Data

19150324

Source:http://linkedlifedata.com/resource/pubmed/id/19150324

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2009-2-16
pubmed:abstractText
Although various nonviral transfection methods are available, cell toxicity, low transfection efficiency, and high cost remain hurdles for in vitro gene delivery in cultured primary endothelial cells. Recently, unprecedented transfection efficiency for primary endothelial cells has been achieved due to the newly developed nucleofection technology that uses a combination of novel electroporation condition and specific buffer components that stabilize the cells in the electrical field. Despite superior transfection efficiency and cell viability, high cost of the technology has discouraged cardiovascular researchers from liberally adopting this new technology. Here we report that a phosphate-buffered saline (PBS)-based nucleofection method can be used for efficient gene delivery into primary endothelial cells and other types of cells. Comparative analyses of transfection efficiency and cell viability for primary arterial, venous, microvascular, and lymphatic endothelial cells were performed using PBS. Compared with the commercial buffers, PBS can support equally remarkable nucleofection efficiency to both primary and nonprimary cells. Moreover, PBS-mediated nucleofection of small interfering RNA (siRNA) showed more than 90% knockdown of the expression of target genes in primary endothelial cells. We demonstrate that PBS can be an unprecedented economical alternative to the high-cost buffers or nucleofection of various primary and nonprimary cells.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-10071611, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-11318112, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-11687237, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-12547715, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-12609550, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-12889809, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-15121163, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-15810891, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-17085816, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-7664017, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-7669041, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-7803234, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-9142319, http://linkedlifedata.com/resource/pubmed/commentcorrection/19150324-9415297
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
1096-0309
pubmed:author
pubmed:issnType
Electronic
pubmed:day
15
pubmed:volume
386
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
251-5
pubmed:dateRevised
2011-9-26
pubmed:meshHeading
pubmed:year
2009
pubmed:articleTitle
Phosphate-buffered saline-based nucleofection of primary endothelial cells.
pubmed:affiliation
Department of Surgery and Biochemistry & Molecular Biology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, 1450 Biggy St. NRT6501, M/C9601, Los Angeles, CA 90033, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Evaluation Studies