title: Magnetofection Mediated Transient NANOG Overexpression Enhances Proliferation and Myogenic Differentiation of Human Hair Follicle Derived Mesenchymal Stem Cells.
abstract: We used magnetofection (MF) to achieve high transfection efficiency into human mesenchymal stem cells (MSCs). A custom-made magnet array, matching well-to-well to a 24-well plate, was generated and characterized. Theoretical predictions of magnetic force distribution within each well demonstrated that there was no magnetic field interference among magnets in adjacent wells. An optimized protocol for efficient gene delivery to human hair follicle derived MSCs (hHF-MSCs) was established using an egfp-encoding plasmid, reaching approximately ∼50% transfection efficiency without significant cytotoxicity. Then we applied the optimized MF protocol to express the pluripotency-associated transcription factor NANOG, which was previously shown to reverse the effects of organismal aging on MSC proliferation and myogenic differentiation capacity. Indeed, MF-mediated NANOG delivery increased proliferation and enhanced the differentiation of hHF-MSCs into smooth muscle cells (SMCs). Collectively, our results show that MF can achieve high levels of gene delivery to MSCs and, therefore, may be employed to moderate or reverse the effects of cellular senescence or reprogram cells to the pluripotent state without permanent genetic modification.
authors: Seoyoung
authors: S
authors: Son
authors: Mao-Shih
authors: MS
authors: Liang
authors: Pedro
authors: P
authors: Lei
authors: Xiaozheng
authors: X
authors: Xue
authors: Edward P
authors: EP
authors: Furlani
authors: ∥Center of Excellence in Bioinformatics and Life Sciences, Buffalo, New York 14203, United States.
authors: Stelios T
authors: ST
authors: Andreadis
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D018859:Hair Follicle / Q000166:cytology*; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D060526:Magnetic Fields; D008280:Magnetics / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D024510:Muscle Development; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D014162:Transfection / Q000379:methods*; D015854:Up-Regulation
chemical_list: D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D004247:DNA