Hi Jonathan,

I'm working with serum-starvation induced autophagy in glial cells and in this cell type 6 hours of serum starvation is more than enough to induce autophagy without killing the cells. In your case, I don't know what is the optimal time to induce autophagy without killing the cells. I would recommend you one of the two options: take a look in the bibliography to see which time is used by the groups who work with monocytes (I suppose you have already do that) or make some experiments in which you make a time course of serum starvation (from 1 to 24 hours for example) and by western-blot check the amount of LC3-II protein and by simple imaging of the cells chech their status.

Hope it helps

Ángel Gaudioso

Thanks a lot for the help. From what I've seen in the literature 6 hours seems about the right amount. I'm going to starve for different amounts of time and see which time is best. Thanks again!

Your goal is to induce autophagy in monocytes or serum starve cells or both?

To induce autophagy in monocytes through serum starvation

Use B12 supplement from sigma. It help in cell proliferation so that cell death can be reduced.

@ Jonathan Anthony Holbrook

Hi Jonathan,

It will be great to know about your experiment. I study autophagy, but I will induce my cells with some pharmaceutical agents. Before incubation, I want to remove serum from medium to reduces interference between growth facts in the FBS and my substance.

Best,

Shirin

Hi Jonathan,

Do you work with primary monocytes?

For how many hours they survive without serum?

Which media do you use?

Serum starvation may increase autophgay flux; however, the use of amino acid deprivation better works. For this, you can use EBSS media. I would also suggest considering shorter times (between 30 min and 6 h).