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Efficient non-viral transfection of THP-1 cells
Authors:Michael Schnoor  Insa Buers  Anika Sietmann  Oliver Hofnagel  Stefan Lorkowski
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
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.
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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