Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation.
Recently, femtosecond laser pulses have been utilized for the targeted introduction of genetic matter into mammalian cells. This rapidly expanding and developing novel optical technique using a tightly focused laser light beam is called phototransfection. Extending previous studies [Stevenson et al., Opt. Express 14, 7125-7133 (2006)], we show that femtosecond lasers can be used to phototransfect a range of different cell lines, and specifically that this novel technology can also transfect mouse embryonic stem cell colonies with approximately 25% efficiency. Notably, we show the ability of differentiating these cells into the extraembryonic endoderm using phototransfection. Furthermore, we present two new findings aimed at optimizing the phototransfection method and improving applicability: first, the influence of the cell passage number on the transfection efficiency is explored and, second, the ability to enhance the transfection efficiency via whole culture treatments. Our results should encourage wider uptake of this methodology.
Mthunzi
Patience
P
University of St. Andrews, School of Physics and Astronomy, North Haugh, St. Andrews, Fife, Scotland, United Kingdom.
Dholakia
Kishan
K
Gunn-Moore
Frank
F
D000818:Animals; D002454:Cell Differentiation / Q000528:radiation effects; D002478:Cells, Cultured; D004247:DNA / Q000493:pharmacokinetics*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology / Q000528:radiation effects*; D007834:Lasers; D051379:Mice; D014162:Transfection / Q000379:methods*
D004247:DNA