How long must HeLa cells be serum starved to induce autophagy? I want to induce autophagy but want a ball park of an idea as to how long they must be placed in serum free media before I can see effects.
If the goal is to induce autophagy in HeLa cells, it could be induced by incubating cultures in HBSS for 30-45 min. However, the induction in the absence of serum could take at least two days or more (depending upon the extent of the removal of growth factors and the origin of cells).
Please note, I have not work on autophagy. But from the practical experience for working with steroids I am telling you that when I put HUVEC in M199 medium without any serum, growth factors, around 30 % of the cells died after 18 hours, around 50% after 24 hours and almost 100% after 48 hours. Almost similar pattern has happened in case of MCF-7 cells in MEM medium. This indicates that you cannot keep the cells free of any serum for two days as indicated above. I totally disagree of that portion of the above answer. Also cell cycle will stop if the medium is completely free of serum and other growth factors.
Please keep in mind that malignant cells often show the secretion of growth factors in the autocrine manner after chronic serum starvation. In the case of HeLa cells, I have experienced the preliminary result that the activation AMPK signal is lost and the paradoxical mTOR activation was observed.
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