title: A Cell Electrofusion Chip for Somatic Cells Reprogramming.
abstract: Cell fusion is a potent approach to explore the mechanisms of somatic cells reprogramming. However, previous fusion methods, such as polyethylene glycol (PEG) mediated cell fusion, are often limited by poor fusion yields. In this study, we developed a simplified cell electrofusion chip, which was based on a micro-cavity/ discrete microelectrode structure to improve the fusion efficiency and to reduce multi-cell electrofusion. Using this chip, we could efficiently fuse NIH3T3 cells and mouse embryonic stem cells (mESCs) to induce somatic cells reprogramming. We also found that fused cells demethylated gradually and 5-hydroxymethylcytosine (5hmC) was involved in the demethylation during the reprogramming. Thus, the cell electrofusion chip would facilitate reprogramming mechanisms research by improving efficiency of cell fusion and reducing workloads.
authors.affiliation: Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, P.R. China; Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.affiliation: Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, P.R. China; Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Qu
authors.affiliation: Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China; Bioengineering College of Chongqing University, Chongqing, P.R. China.
authors.forename: Ning
authors.initials: N
authors.lastname: Hu
authors.affiliation: Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, P.R. China; Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China.
authors.forename: Yuxiao
authors.initials: Y
authors.lastname: Zeng
authors.affiliation: Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China; Bioengineering College of Chongqing University, Chongqing, P.R. China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, P.R. China; Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China.
authors.forename: Haiwei
authors.initials: H
authors.lastname: Xu
authors.affiliation: Southwest Eye Hospital, Southwest Hospital, Third Military Medical University, Chongqing, P.R. China; Key Lab of Visual Damage and Regeneration & Restoration of Chongqing, Chongqing, P.R. China.
authors.forename: Zheng Qin
authors.initials: ZQ
authors.lastname: Yin
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D065150:Cellular Reprogramming; D003596:Cytosine / Q000031:analogs & derivatives / Q000378:metabolism; D004247:DNA / Q000032:analysis; D019175:DNA Methylation; D018274:Electroporation / Q000295:instrumentation*; D018398:Homeodomain Proteins / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008819:Mice, Nude; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D041681:NIH 3T3 Cells; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA