Document Detail


Improved protocol for efficient nonviral transfection of premature THP-1 macrophages.
MedLine Citation:
PMID:  21536764     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The human monocytic leukemia cell line THP-1 is a widely used model for investigating monocyte and macrophage biology. Successful transfection of THP-1 monocytes with subsequent phorbol 12-myristate 13-acetate (PMA)-induced differentiation into macrophages is not a trivial matter, because according to previous transfection protocols, cell viability is lost almost completely within 24 h of PMA treatment following transfection. This protocol constitutes an optimized version of a previously published protocol by our group. It describes a procedure for transfecting premature THP-1 macrophages, which subsequently can be further differentiated into mature macrophages by PMA without a loss of cell viability. Transfection of THP-1 cells with plasmids or small interfering RNA (siRNA) is achieved by electroporation using the Lonza Nucleofector technology (Basel, Switzerland). This technique allows for the efficient nonviral delivery of plasmids, DNA, RNA, or siRNA into primary cells or cell lines even if the cells are not or are only slowly proliferating. Such cells are usually rather difficult to transfect by nonviral approaches. This means that only viral approaches would be left, which are expensive and labor-intensive and require laboratories complying with the respective safety regulations. The protocol described here is an efficient and convenient alternative.
Authors:
Marten B Maess; Insa Buers; Horst Robenek; Stefan Lorkowski
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-05-01
Journal Detail:
Title:  Cold Spring Harbor protocols     Volume:  2011     ISSN:  1559-6095     ISO Abbreviation:  Cold Spring Harb Protoc     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-05-03     Completed Date:  2011-08-09     Revised Date:  2013-05-20    
Medline Journal Info:
Nlm Unique ID:  101524530     Medline TA:  Cold Spring Harb Protoc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  pdb.prot5612     Citation Subset:  IM    
Affiliation:
Institute of Nutrition, Friedrich Schiller University Jena, 07743 Jena, Germany.
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MeSH Terms
Descriptor/Qualifier:
Cell Differentiation / drug effects
Cell Line, Tumor
Cell Survival / drug effects
Electroporation / methods
Humans
Macrophages / physiology*
Plasmids / metabolism
RNA, Small Interfering / metabolism
Tetradecanoylphorbol Acetate / metabolism
Transfection / methods*
Chemical
Reg. No./Substance:
0/RNA, Small Interfering; 16561-29-8/Tetradecanoylphorbol Acetate

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


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