title: Generation of murine induced pluripotent stem cells by using high-density distributed electrodes network.
abstract: This study reports a robust method of gene transfection in a murine primary cell model by using a high-density electrodes network (HDEN). By demonstrating high cell viability after gene transfection and successful expression of transgenes including fluorescent proteins, the HDEN device shows great promise as a solution in which reprogramming efficiency using non-viral induction for generation of murine induced pluripotent stem cells (iPSCs) is optimized. High and steady transgene expression levels in host cells of iPSCs can be demonstrated using this method. Moreover, the HDEN device achieved successful gene transfection with a low voltage of less than 180 V while requiring relatively low cell numbers (less than 1.5 × 10(4) cells). The results are comparable to current conventional methods, demonstrating a reasonable fluorescent-plasmid transfection rate (42.4% in single transfection and 24.5% in triple transfection) and high cell viability of over 95%. The gene expression levels of each iPSC factor was measured to be over 10-fold higher than that reported in previous studies using a single mouse embryonic fibroblast cell. Our results demonstrate that the generation of iPSCs using HDEN transfection of plasmid DNA may be a feasible and safe alternative to using viral transfection methods in the near future.
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University , Hsinchu, Taiwan.
authors.forename: Ming-Yu
authors.initials: MY
authors.lastname: Lu
authors.affiliation: National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Institute of Microelectronics, Peking University , Beijing, China.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute , Hsinchu, Taiwan.
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Regenerative Medicine Research Group, Institute of Cellular and System Medicine, National Health Research Institutes , Zhunan, Taiwan.
authors.forename: B
authors.initials: B
authors.lastname: Linju Yen
authors.forename: Gwo-Bin
authors.initials: GB
authors.lastname: Lee