Gene transfer via nucleofection into adult and embryonic stem cells.
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.
Lakshmipathy
Uma
U
Stem Cell Institute, University of Minnesota, Minneapolis, USA.
Buckley
Shannon
S
Verfaillie
Catherine
C
D053687:Adult Stem Cells / Q000378:metabolism*; D000818:Animals; D001853:Bone Marrow / Q000502:physiology*; D018274:Electroporation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D053652:Nuclear Transfer Techniques; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D049452:Green Fluorescent Proteins