Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells.

Abstract:

: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.

journal_name

J Neurosci Methods

authors

Kim YC,Shim JW,Oh YJ,Son H,Lee YS,Lee SH

doi

10.1016/s0165-0270(02)00090-0

subject

Has Abstract

pub_date

2002-06-30 00:00:00

pages

153-8

issue

2

eissn

0165-0270

issn

1872-678X

pii

S0165027002000900

journal_volume

117

pub_type

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