I am currently doing some work about starvation-induced autophagy.
I treated the cells with EBSS medium without FBS for 4 h to induce autophagy. The morphology of the cells got strange. And when I changed the medium to normal DMEM, I found that a lot of cells was washed away.
I am wondering if I could shorten the period of time to induce autophagy? I have used Hela and MCF7, these two cell lines. But the results were the same. Thanks, everybody.
Hela and MCF7 won't die normally under 4h starvation (see multiple papers from the Kroemer lab). Most likely, the problem is that you have Ca2-free EBSS, which detaches the cells. Make sure that your EBSS contains Ca2 as a first step
Hi Tianyi
I am agree with you.. Try starvation in different periods of time toget the optimal time for inducing autophagy in your cell lines.
Research = trial and error
If you are rinsing cells with PBS before changing to EBSS, incubation should be 20 min to 40 min (maximum).
Hela and MCF7 won't die normally under 4h starvation (see multiple papers from the Kroemer lab). Most likely, the problem is that you have Ca2-free EBSS, which detaches the cells. Make sure that your EBSS contains Ca2 as a first step
Thanks for reply and help from you all. Particularly, Dr. Galluzzi I think my case is more like what you said. The cells may be detached from the plate. But the medium I used is from Thermo, 24010043 which contains 200 mg/L Ca2+. I wonder if this is enough to attach the cells to the plate? Thank you.
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