Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells.
Inefficiency in primary neural stem cell transfection is a major obstacle in gene expression research aimed at determining the mechanisms underlying brain development. Following our initial finding that liposome-mediated transfection procedures are fairly toxic to neural stem cells, we further examined whether transfection efficiency could be improved by preventing cells from undergoing apoptosis. Transfection efficiencies were markedly enhanced by co-transfection of cells with prototypic anti-apoptotic genes, such as bcl-2 and bcl-xL, and supplementing the culture medium with B27 Supplement. This combination of anti-apoptotic gene co-transfection and B27 Supplement resulted in approximately 5% transfection efficiency of primary neural stem cells, compared to less than 0.2% in control transfections. Therefore, this procedure and other similar approaches employed to enhance the efficiency of transfecting neural stem cells may facilitate the understanding of mechanisms underlying self-renewal of neural stem cells and their differentiation into various cell lineages.
Kim
You Chan
YC
Department of Biochemistry, College of Medicine, Hanyang University, 133-791, Seoul, South Korea.
Shim
Jae Won
JW
Oh
Young J
YJ
Son
Hyeon
H
Lee
Yong Sung
YS
Lee
Sang Hun
SH
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D003470:Culture Media / Q000494:pharmacology; D018076:DNA, Complementary; D005333:Fetus / Q000166:cytology; D049452:Green Fluorescent Proteins; D020287:In Situ Nick-End Labeling; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010957:Plasmids / Q000493:pharmacokinetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D051381:Rats; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D014162:Transfection / Q000379:methods*; D051020:bcl-X Protein
C494813:Bcl2l1 protein, rat; D003470:Culture Media; D018076:DNA, Complementary; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051020:bcl-X Protein; D049452:Green Fluorescent Proteins