title: Optimization of the transfection of human THP-1 macrophages by application of Nunc UpCell technology.
abstract: We have established an electroporation protocol for transfection of premature adherent human THP-1 macrophages using Lonza Nucleofector technology. For efficient electroporation, detachment of adherent cells is necessary. We tested the Nunc UpCell product line of Thermo Fisher Scientific, which achieves detachment by a change of ambient temperature, as an alternative to enzymatic detachment. Here we present data verifying proper cell morphology and vitality and high transfection efficiency for macrophages cultured on UpCell plates. Appropriate macrophage behavior was confirmed by measuring markers of macrophage differentiation and polarization by reverse transcription quantitative polymerase chain reaction (RT-qPCR). In conclusion, Nunc UpCell materials are a viable alternative to enzymatic detachment.
authors: Institute of Nutrition, Friedrich Schiller University Jena, 07743 Jena, Germany.
authors: Marten B
authors: MB
authors: Maeß
authors: Institute of Nutrition, Friedrich Schiller University Jena, 07743 Jena, Germany.
authors: Andrea-Anneliese
authors: AA
authors: Keller
authors: Department of Molecular Hemostaseology, Jena University Hospital, 07743 Jena, Germany.
authors: Knut
authors: K
authors: Rennert
authors: Department of Molecular Hemostaseology, Jena University Hospital, 07743 Jena, Germany; Integrated Research and Treatment Center, Center for Sepsis Control and Care (CSCC), Jena University Hospital, 07747 Jena, Germany.
authors: Alexander
authors: A
authors: Mosig
authors: Institute of Nutrition, Friedrich Schiller University Jena, 07743 Jena, Germany. Electronic address: stefan.lorkowski@uni-jena.de.
authors: Stefan
authors: S
authors: Lorkowski
mesh_terms: D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering
keywords: Detachment; THP-1 macrophages; Transfection; siRNA