Efficient generation of virus-free iPS cells using liposomal magnetofection.
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.
Miraebio Research Institute, Mirae Biotech, Seoul, Korea.
Hyo Young
HY
Park
Eun Hyung
EH
Noh
Hyung-Min
HM
Chung
Man-Jong
MJ
Kang
Eun Young
EY
Kim
Se Pill
SP
Park
D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005821:Genetic Techniques; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D008280:Magnetics; D051379:Mice; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology; D013724:Teratoma / Q000378:metabolism; D013997:Time Factors; D019076:Transgenes
D008081:Liposomes; D004247:DNA; D007770:L-Lactate Dehydrogenase