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.
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Acknowledgements
We would like to thank Dr. Raymond Ching-Bong Wong and Marek Mandau for helpful comments on this manuscript. This work was supported by funding from the National Institutes of Health (HD149488; HD47675), the California Institute for Regenerative Medicine (RC1-00110), and by institutional funds from the University of California, Irvine to P.J.D.; by a CIRM Pre-doctoral Training Fellowship (T1-00008) and by the Cellular and Molecular Biosciences Graduate Program at the University of California, Irvine to H.F.; by the Human Genetics and Molecular Biology Training Program (NIH 2T32GM07814) at Johns Hopkins University School of Medicine to K.A.H.E.; and by institutional funds from the University of California, Irvine to L.F.L.
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© 2011 Springer Science+Business Media, LLC
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Fong, H., Elliott, K.A.H., Lock, L.F., Donovan, P.J. (2011). Nucleofection of Human Embryonic Stem Cells. In: Schwartz, P., Wesselschmidt, R. (eds) Human Pluripotent Stem Cells. Methods in Molecular Biology, vol 767. Humana Press. https://doi.org/10.1007/978-1-61779-201-4_24
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DOI: https://doi.org/10.1007/978-1-61779-201-4_24
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Online ISBN: 978-1-61779-201-4
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