Expression of enhanced green fluorescent protein gene in PC12 cells

Aim: To study expression of enhanced GFP gene in PC12 cells and its application. Methods: PC12 cells were transfected by enhanced GFP gene in liposome-mediated gene transfer, GFP-expression and fluorescent intensity were detected under fluorescent microscopy. The PC12/GFP cells after anoxia-serium deprivation/reperfusion were stained by PI and we tried to sort the GFP positive survival cells and determine the percentage of them. Results: Almost all of the cells expressed high-level GFP after being selected by G418, and there was no significant difference between fluorescent intensity of the 5th generation PC12/GFP cells and that of the 40th. Four kinds of cells including GFP( + ) / PI( - ) ,GFP( + ) / PI( + ) ,GFP( - ) / PI( - ) ,GFP( - ) / PI( + ) were accurately sorted by flow cytometry. Conclusions: The PC12/ GFP cell line is a stable, high-level GFP-expression cell line, and it can be served as an ideal control group for the study of neuroprotection after anoxia-serium deprivation/reperfusion.