title: Innovative strategy for microRNA delivery in human mesenchymal stem cells via magnetic nanoparticles.
abstract: Bone marrow derived human mesenchymal stem cells (hMSCs) show promising potential in regeneration of defective tissue. Recently, gene silencing strategies using microRNAs (miR) emerged with the aim to expand the therapeutic potential of hMSCs. However, researchers are still searching for effective miR delivery methods for clinical applications. Therefore, we aimed to develop a technique to efficiently deliver miR into hMSCs with the help of a magnetic non-viral vector based on cationic polymer polyethylenimine (PEI) bound to iron oxide magnetic nanoparticles (MNP). We tested different magnetic complex compositions and determined uptake efficiency and cytotoxicity by flow cytometry. Additionally, we monitored the release, processing and functionality of delivered miR-335 with confocal laser scanning microscopy, real-time PCR and live cell imaging, respectively. On this basis, we established parameters for construction of magnetic non-viral vectors with optimized uptake efficiency (~75%) and moderate cytotoxicity in hMSCs. Furthermore, we observed a better transfection performance of magnetic complexes compared to PEI complexes 72 h after transfection. We conclude that MNP-mediated transfection provides a long term effect beneficial for successful genetic modification of stem cells. Hence, our findings may become of great importance for future in vivo applications.
authors.affiliation: Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, University of Rostock, Schillingallee 35, 18057 Rostock, Germany. gustav.steinhoff@med.uni-rostock.de.
authors.forename: Anna
authors.initials: A
authors.lastname: Schade
authors.forename: Evgenya
authors.initials: E
authors.lastname: Delyagina
authors.forename: Dorothee
authors.initials: D
authors.lastname: Scharfenberg
authors.forename: Anna
authors.initials: A
authors.lastname: Skorska
authors.forename: Cornelia
authors.initials: C
authors.lastname: Lux
authors.forename: Robert
authors.initials: R
authors.lastname: David
authors.forename: Gustav
authors.initials: G
authors.lastname: Steinhoff
mesh_terms: D002465:Cell Movement; D055785:Gene Knockdown Techniques; D018014:Gene Transfer Techniques; D006801:Humans; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
chemical_list: D058185:Magnetite Nanoparticles; D035683:MicroRNAs