In recent months, I have tried to establish an in vitro immortalized mouse model of high glucose damage to observe the damage of podocytes and mitochondria by high glucose in vitro, but unfortunately, no obvious damage was observed (cell morphology, CCK8 And JC-1 are not significantly different from the control group& High glucose group).

CCK-8: I seeded 1 × 10 ^ 4 / well, serum-free for 24h after night, and then add low- or high-glucose 1640 medium containing 10% FBS?

Is my cell density too high? Or should I not add 10% FBS to the high-sugar medium? (But my total research time is up to 72h. Will, cells do not die without FBS? After reading a lot of paper, many researchers seem to avoid mentioning whether to add FBS after serum starvation)

Do you have any experience in establishing high-glucose injury models of in vitro immortalized podocytes or other cells? I hope to get your share, thanks!