Efficient non-viral transfection of THP-1 cells.
Macrophages are an important part of the cellular immune system and play a key role during immune responses. Thus, macrophages are interesting targets in basic and clinical research. Primary monocytes or monocyte-derived macrophages do not proliferate on a suitable scale so that their use for functional studies in vitro is limited. Immortal proliferating cell lines, such as the human THP-1 monocytic leukemia cell line, are therefore often used instead of primary cells. Transfection is a useful tool to study the function of gene products, but transfection of THP-1 monocytes and pre-differentiated THP-1 macrophages with subsequent differentiation into mature THP-1 macrophages using phorbol esters is usually accompanied by a progressive loss of cell viability. In this study, we describe a simple and rapid approach for efficient transfection of THP-1 monocytes and pre-differentiated THP-1 macrophages using a modified Nucleofection-based approach. The protocol maintains cell viability and functionality, thus allowing efficient transfection of THP-1 cells combined with subsequent differentiation of transfected THP-1 cells into mature macrophages.
Schnoor
Michael
M
Leibniz Institute of Arteriosclerosis Research, Domagkstrasse 3, 48149 Münster, Germany.
Buers
Insa
I
Sietmann
Anika
A
Brodde
Martin F
MF
Hofnagel
Oliver
O
Robenek
Horst
H
Lorkowski
Stefan
S
D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002470:Cell Survival; D015870:Gene Expression; D006801:Humans; D008264:Macrophages / Q000166:cytology*; D009000:Monocytes / Q000166:cytology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D012333:RNA, Messenger