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PMID: 2786626 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Enhanced transfection efficiency and improved cell survival after electroporation of G2/M-synchronized cells and treatment with sodium butyrate.

Nucleic acids research ·Vol. 17 ·No. 10 ·1989-05-25 ·Pages 3959-71

Goldstein S, Fordis CM, Howard BH

Abstract

To achieve high transfection efficiency in human fibroblasts with good preservation of proliferative capacity we developed an electroporation procedure that combines two distinct modalities: use of recipient cells synchronized in the late G2/mitotic phase of the cell cycle and treatment of cells post-electroporation with 5 mM butyrate. This combination enabled reduction of plasmid DNA concentration and electroporation voltage, both associated with cytotoxicity, while greatly enhancing transfection efficiencies. Although the method was primarily developed for transient expression it was also found to improve stable expression. This procedure should have wide applicability, particularly in studies seeking to identify DNA sequences that lead to inhibition of DNA synthesis and proliferation in human fibroblasts and other cells refractory to transfection.

MeSH Terms
Butyrates/pharmacology Butyric Acid Cell Cycle/drug effects Cell Membrane Permeability Cell Survival/drug effects Electrophysiology/methods Fibroblasts/cytology,drug effects Genes Humans Macromolecular Substances Plasmids Receptors, Interleukin-2/genetics Transfection
Chemicals
Butyrates Macromolecular Substances Receptors, Interleukin-2 Butyric Acid
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Goldstein S
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892.
Fordis C M
Howard B H
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.10.3959
Published
1989-05-25
Pages
3959-71
Language
English
Region
England
NLM ID
0411011
PMCID
PMC317872
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.10.3959 PMC Full Text View journal details
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