Protocol

Stem Cell Assays

Volume 407 of the series Methods in Molecular Biology™ pp 115-126

Gene Transfer Via Nucleofection Into Adult and Embryonic Stem Cells

  • Uma LakshmipathyAffiliated withStem Cell Institute, University of MinnesotaStem Cell Institute, Invitrogen Corporation
  • , Shannon BuckleyAffiliated withStem Cell Institute, University of Minnesota
  • , Catherine VerfaillieAffiliated withStem Cell Institute, University of Minnesota

* Final gross prices may vary according to local VAT.

Get Access

Summary

The use of embryonic and adult stem cells as therapeutic agents is gaining momentum. A major impediment in the use of stem cells for genetic disorders is their ability to undergo genetic modification. The recognition of various site-specific integration methods open up a new avenue for gene therapy in stem cells. However, this necessitates efficient delivery of DNA molecule into cells. Most commercially used liposome-mediated transfection reagents are toxic or work poorly with stem cells. Electroporation, while effective in transfecting stem cells, is rather harsh and leads to excessive cell death. Nucleofection, a technology by Amaxa, uses a combination of electric pulse in an appropriate media, which decreases the toxicity and promotes efficient transfection of stem cells. Various types of adult and embryonic stem cells can be successfully transfected using this method, as described in this chapter.

Keywords

Mouse bone marrow Embryonic stem cells Adult stem cells Transfection Nucleofection