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Application of pulsed-magnetic field enhances non-viral gene delivery in primary cells from different origins
Authors:Sarah W Kamau Chapman  Paul O Hassa  Sabine Koch-Schneidemann  Brigitte von Rechenberg  Margarethe Hofmann-Amtenbrink  Benedikt Steitz  Alke Petri-Fink  Heinrich Hofmann  Michael O Hottiger
Institution:1. Institute of Veterinary Biochemistry and Molecular Biology, University of Zurich, Winterthurerstr. 190, 8057 Zurich, Switzerland;2. European Molecular Biology Laboratory (EMBL) Heidelberg, Meyerhofstrasse 1, 69117 Heidelberg, Germany;3. Musculoskeletal Research Unit, Equine Hospital, Vetsuisse Faculty Zurich, University of Zurich, Winterthurerstr. 260, 8057 Zurich, Switzerland;4. MatSearch, Chemin Jean Pavillard 14, 1009 Pully, Switzerland;5. Laboratory of Powder Technology, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland
Abstract:Primary cell lines are more difficult to transfect when compared to immortalized/transformed cell lines, and hence new techniques are required to enhance the transfection efficiency in these cells. We isolated and established primary cultures of synoviocytes, chondrocytes, osteoblasts, melanocytes, macrophages, lung fibroblasts, and embryonic fibroblasts. These cells differed in several properties, and hence were a good representative sample of cells that would be targeted for expression and delivery of therapeutic genes in vivo. The efficiency of gene delivery in all these cells was enhanced using polyethylenimine-coated polyMAG magnetic nanoparticles, and the rates (17–84.2%) surpassed those previously achieved using other methods, especially in cells that are difficult to transfect. The application of permanent and pulsating magnetic fields significantly enhanced the transfection efficiencies in synoviocytes, chondrocytes, osteoblasts, melanocytes and lung fibroblasts, within 5 min of exposure to these magnetic fields. This is an added advantage for future in vivo applications, where rapid gene delivery is required before systemic clearance or filtration of the gene vectors occurs.
Keywords:Gene delivery  Primary cell  Magnetic nanoparticle  Polyethylenimine  Magnet
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