Efficient generation of virus-free iPS cells using liposomal magnetofection

• Published in: PloS one Vol. 7; no. 9; p. e45812
• Main Authors: Park, Hyo Young, Noh, Eun Hyung, Chung, Hyung-Min, Kang, Man-Jong, Kim, Eun Young, Park, Se Pill
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.09.2012; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Biocompatibility; Biology; Biotechnology; Cell Culture Techniques; Cell lines; Cells, Cultured; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - metabolism; Efficiency; Electron microscopy; Embryo fibroblasts; Fibroblasts; Gene expression; Genetic engineering; Genetic Techniques; Induced Pluripotent Stem Cells - cytology; Integration; L-Lactate dehydrogenase; L-Lactate Dehydrogenase - metabolism; Lactate dehydrogenase; Lactic acid; Liposomes - metabolism; Magnetic fields; Magnetics; Materials Science; Medical research; Mice; Microscopy, Electron, Transmission - methods; Models, Genetic; Morphology; Nanoparticles; Nanotechnology; Plasmids; Plasmids - metabolism; Pluripotency; Regenerative medicine; Smooth muscle; Stem cell research; Stem cells; Stem Cells - cytology; Teratoma; Teratoma - metabolism; Time Factors; Transfection; Transgenes; Transmission electron microscopy; Viral infections; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0045812

The generation of induced pluripotent stem (iPS) cells is a powerful tool in regenerative medicine, and advances in nanotechnology clearly have great potential to enhance stem cell research. Here, we introduce a liposomal magnetofection (LMF) method for iPS cell generation. Efficient conditions for generating virus-free iPS cells from mouse embryonic fibroblast (MEF) cells were determined through the use of different concentrations of CombiMag nanoparticle-DNA(pCX-OKS-2A and pCX-cMyc)-lipoplexes and either one or two cycles of the LMF procedure. The cells were prepared in a short reprogramming time period (≤ 8 days, 0.032-0.040%). Among the seven LMF-iPS cell lines examined, two were confirmed to be integration-free, and an integration-free LMF-iPS cell line was produced under the least toxic conditions (single LMF cycle with a half-dose of plasmid). This cell line also displayed in vitro/in vivo pluripotency, including teratoma formation and chimeric mouse production. In addition, the safety of CombiMag-DNA lipoplexes for the transfection of MEF cells was confirmed through lactate dehydrogenase activity assay and transmission electron microscopy. These results demonstrated that the LMF method is simple, effective, and safe. LMF may represent a superior technique for the generation of virus-free or integration-free iPS cell lines that could lead to enhanced stem cell therapy in the future.