A NEW TOOL FOR EFFICIENT TRANSFECTION OF DOG AND HUMAN THYROCYTES IN PRIMARY CULTURE

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.