4 replies to this topic

[AnnaSh]

Hi guys!

I'd like to analyze the apoptotic effect of a peptide molecule on cultured cells. I usually use 12 hour serum-starved cells and I apply the compound dissolved in serum-free medium up to 96 hours. I'm afraid that using a serum-free medium during a long  treatment can affect health of cells even in the absence of the compound.
Do you think I'm right when using serum-free medium during treatments? Can you suggest some general rules to conduct treatments in cell cultures? Is absence of serum essential during tests? What compounds contained in standard medium must be avoided in treatment media?

Thank you very much for your replies!

[bob1]

Serum starvation causes the cells to stop cycling (at G1 if I recall correctly), this may affect how the cells respond to the drugs, especially if the peptide binds to something that is part of the cell cycle such as microtubule formation.

[AnnaSh]

Yes, you're right, but serum starvation (for 12 hr at least) is essential to synchronize cell cycles before administering treatments. My doubts concern the lenght of treatments, since I Know that a prolonged serum depletion can induce apoptosis. You this I have to resuspend my compounds in serum-containing medium?

Thanks a lot!

[bob1]

I would try and keep the cell conditions as "normal" as possible.  You should be able to dissolve the compounds in a serum free medium and then dilute in serum containing medium.

If you are using a commonly used cell line such as 293, A549, MCF-7, i would try and see if the cells function normally with reduced serum at a range of concentrations and then use the minimum possible.

[AnnaSh]

I'll try to keep the serum content as low as possible, but won't eliminate it completely (even if it's a diffuse custom (!) I haven't ever really understood...).

Thanks a lot for your suggestion!