I'm using HepG2 cells (and have AML12 available). I've tried serum-free media and glucose-free media, but it doesn't quite mimic the hepatic fasting response in mice, at least according to key gene expression changes (gluconeogenesis and fatty acid oxidation genes).
This difference is somewhat expected. Try to add 2 uM of Oleanolic Acid to your starvation medium and check by qPCR if the key gene expression changes are similar to results obtained in the fasting mice. In starving mammals, the primary stress factor is not the insufficient energy it is toxicity from the products of fatty acids katabolism in the body, adding Oleanolic acid will mimic this factor.
Duration of treatment is also important. In my own experience, for some cells lines 72 h (serum-free or glucose-free medium) is a minimum setting for detection of significant changes/differences between starving and normal conditions. Starvation activates autophagy and cells can keep going for 48h and longer with no detectable changes.
What if you seeded to lowish culture and just didnt replace medium as they grew, nutrient will get low, thus starvation condition. . ?
Best
- Badges
- Science topic