N. Uyttersprot et al., A NEW TOOL FOR EFFICIENT TRANSFECTION OF DOG AND HUMAN THYROCYTES IN PRIMARY CULTURE, Molecular and cellular endocrinology, 142(1-2), 1998, pp. 35-39
The introduction of exogenous DNA into mammalian cells is commonly use
d to study the functions of gene products. However cells in primary cu
lture are usually refractory to most transfection systems. Here we inv
estigated the ability of a new lipid formulation, FuGENE(TM) 6 transfe
ction reagent, to promote DNA uptake into dog and human thyroid cells
in primary culture. Gene transfer was monitored by the expression of a
Green Fluorescent Protein (GFP) reporter gene. We report that FuGENE
6 is particularly suited for the transfection of thyroid cells and doe
s not interfere with their normal growth. Optimization of the experime
ntal conditions, such as DNA amount, DNA/lipid ratio, cell density and
incubation with the transfection mixture, was achieved by evaluating
the percentage of GFP-expressing cells by FAGS analysis. FuGENE 6 allo
wed us to obtain 8-15% thyrocytes expressing the reporter gene which r
epresents an efficiency 100-fold superior to other transfection method
s. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.