title: Nucleofection of human embryonic stem cells.
abstract: The ability to realize the full potential of human pluripotent stem cells (hPSCs) as tools for -understanding human development and advancing the field of regenerative medicine is dependent on efficient methods to genetically manipulate these cells. There are several methods for introducing foreign DNA into cells such as electroporation, lipid-based transfection technology, and viral transduction. We describe here a method to transfect human embryonic stem cells (hESCs) using nucleofection technology. This unique method uses the Nucleofector II Device that combines the use of a cell type-specific Nucleofector Solution and preprogrammed electrical parameters to efficiently deliver DNA into the cell nucleus. The use of this technology allows high-efficiency transfer of nucleic acids into hESCs enabling both transient and stable manipulation of gene expression in these cells.
authors.affiliation: Department of Biological Chemistry, Sue and Bill Gross Stem Cell Research Center, School of Medicine, University of California, Irvine, Irvine, CA, USA.
authors.forename: Helen
authors.initials: H
authors.lastname: Fong
authors.forename: K A Hohenstein
authors.initials: KA
authors.lastname: Elliott
authors.forename: Leslie F
authors.initials: LF
authors.lastname: Lock
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Donovan
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D051379:Mice; D034421:Sus scrofa; D014162:Transfection / Q000379:methods*
chemical_list: D007202:Indicators and Reagents; D049452:Green Fluorescent Proteins