I want to induce apoptosis or oxidative stress by High glucose in HEK293 cells. But the media already contains 4.5g/L of glucose (25mM). How should i approach such experiments?
You need to use media without glucose to prepared desired concentration using sterile glucose solution, e.g. 2.5mM, 11mM and 25mM etc. Incubate your cells using these medias at varying time point as per your experiments. Then measure apoptosis or oxidative stress. You can incubate different concentration at different time point and check the readout. Hope this helps.
In most of the articles which i read regarding high glucose induced toxicity, they showed generation of oxidative stress induced by high glucose which subsequently followed by apoptosis.