700 mg bid

Rat pheochromocytoma cell lines (PC12) were cultured in RPMI-1640 supplemented with 5% FBS and 10% HS, at 37◦C in a humidified atmosphere of 5% CO2 and 95% oxygen. The cells were incubated with Na2S2O4 at a final concentration of 10mM in the sugar and glutamine-free Earle’s solution for 4 h (hypoxia) and the cell solution was then replaced by normal medium without adding glutamine (reoxygenation). Series concentrations of Aceglutamide (0.01, 0.1, 1 and 10 mkM) were added into the culture medium 24 h after the onset of reoxygenation. Control cultures were treated in a similar way except the hypoxic challenge.