Hi!
I have a problem with low glutamate toxicity on non-differentiated PC-12 cells. I use a concentration of 10-20 mM glutamate, add to the culture medium for 24 hours, ~ 20% of the cells die (MTT test). I cultivate in the medium DMEM with 10% FBS.
I think that you are using a higher concentration of glutamate. The conc. of L-glutamine for DMEM is 4 mM (attached).
Adding extra glutamate can inhibit growth/proliferation of cells. See link: https://www.researchgate.net/post/What_is_the_role_of_L-Glutamine_in_media_and_what_happen_if_it_is_inactivated_by_leaving_the_media_overnight_at_37C
Varun Ahuja
I need to achieve neurotoxicity, as I test substances that have a protective effect, but it seems to me that the toxicity is too low
OK Georg..So, I got your question wrong initially. What about the possibility of increasing the conc. of glutamate further?
As far as I understand, concentrations greater than 10 mM (I have tried 20 and even 50 but there is no difference) are not used. I get that there is no dose-dependent effect, I would like to know from those who used the glutamate toxicity model is this true? Maybe I'm not doing the experiment correctly.
My task is to evaluate the protective effect of certain substances in the model of glutamate toxicity on PC-12 cells
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