Efficient non-viral transfection of THP-1 cells

Schnoor M, Buers I, Sietmann A, Brodde MF, Hofnagel O, Robenek H, Lorkowski S.
Source: J Immunol Methods
Publication Date: (2009)
Issue: 344(2): 109-15
Research Area:
Immunotherapy / Hematology
Cells used in publication:
Species: human
Tissue Origin: blood
Macrophage, human
Species: human
Tissue Origin: blood
Nucleofector® I/II/2b

Results: 50% TE and 80% VIA for THP-1 monocytes 40% TE and 75% VIA for THP-1 macrophages Changes to recommended protocol for NuFe II: Human Monocyte Nucleofector Kit instead of Cell Line Nucleofector Kit V. (I suppose the program used was V-001 as in the THP-1 OP) 20% autologous human serum (PAA) instead of 10% fetal bovine serum 2.5*10e6 cells per transfection, and pmaxGFP was reduced to 0.5 µg per cuvette. For gene silencing, 1 µg of siRNA directed against TIP47 or scrambled siRNA (Qiagen, Hilden, Germany) were used. Recovery phase of 4 h after transfection in Human Monocyte Nucleofector Medium supplemented with 20% human serum, 1% non-essential amino acids, 1% sodium pyruvate and 0.1 mg/ml penicillin/streptomycin/L-glutamine at 37 °C and 5% carbon dioxide. Detailed step by- step protocols for the transfection of THP-1 monocytes and pre-matured THP-1 macrophages are provided as online supplemental material.


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.