Efficient non-viral transfection of THP-1 cells |
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Authors: | Michael Schnoor Insa Buers Anika Sietmann Oliver Hofnagel Stefan Lorkowski |
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Affiliation: | a Leibniz Institute of Arteriosclerosis Research, Domagkstraße 3, 48149 Münster, Germany b Emory University, Department of Pathology and Laboratory Medicine, 615 Michael Street, Atlanta, GA, United States c OxProtect, Mendelstraße 11, 48149 Münster, Germany d Institute of Nutrition, Friedrich Schiller University Jena, Dornburger Straße 25, 07743 Jena, Germany |
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Abstract: | 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 invitro 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. |
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Keywords: | ADRP, adipophilin, adipose differentiation-related protein GAPDH, glyceraldehyde-3-phosphate dehydrogenase GFP, green fluorescent protein HKG, housekeeping gene(s) LPS, lipopolysaccharide PAT, protein family comprised of perilipin, adipophilin and TIP47 PI, propidium iodide PMA, phorbol 12-myristate 13-acetate SRP14, 14  kDa signal recognition particle TIP47, 47  kDa MPR tail-interacting protein, mannose 6-phosphate receptor-binding protein 1 (M6PRBP1) TUBA1, α1 tubulin UBE2D2, ubiquitin-conjugating enzyme E2D 2. |
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