Nucleofection of human embryonic stem cells.
Human embryonic stem cells (HESCs) are widely used as a model system for human cell type specification. Genetic modification forms a valuable tool for HESC technology, as it provides the basis for lineage selection, i.e., the purification of a specific cell type after differentiation. Electroporation is an efficient way to transfect HESCs. Nucleofection is an electroporation-based transfection technique which utilizes cell-type-specific buffer solutions and specific electric settings. Customization of these two parameters has been proven to result in highly efficient gene transfer even in hard-to-transfect cells. We can show that nucleofection surpasses conventional electroporation in efficiency and decreases the experimental effort for transfection of HESCs.
Siemen
Henrike
H
Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn, Bonn, Germany.
Nolden
Lars
L
Terstegge
Stefanie
S
Koch
Philipp
P
Brüstle
Oliver
O
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D004274:DNA, Recombinant; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins