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Methods Mol Biol. 2008;423:131-8. doi: 10.1007/978-1-59745-194-9_8.

Nucleofection of human embryonic stem cells.

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1
Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn, Bonn, Germany.

Abstract

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.

PMID:
18370194
DOI:
10.1007/978-1-59745-194-9_8
[Indexed for MEDLINE]
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