Hi all,
I am running a n experiment involving primary rat Schwann cells ( passage 3) and varying dosages of H202 (0mM, 0.1mM, 0.3mM, 0.8mM, 1mM, and 2mM). Schwann cells ( n=2,000) were grown on a 96 well plate for 48 hours in 10% DMEM media with glutamax and forskulin. The varying concentration of H202 ( previously mentioned) was added to media after 48 hours and cells were subsequently grown in 1%FBS media. CellsThe experiment was done in triplicates. The results of my experiment seemed to show an increase in Schwann cell proliferation after CCK-8 assay. Is this to be expected or is it a technical error?
Please see attached for my data results
Cell death (Necrotic mode) might be a more relevant pheromone in this case (High H2O2 concentrations)!
I think that you are in front of a new discovery. You should go deeper into the issue. Even if increased anti-oxidant response in Schwann cells have been described before, your finding is the first one showing an increase in the reproduction of cells. I do not think it is a technical error.
Hi Tomas Koltai , I speculate that my results are due to the nature of the experiment. CCK-8 assay uses the WST-8 protein that illuminates after reacting with NADH. The level of NADH, measured by WST-8 mediated illumination, may have been upregulated as a response to H202 mediated oxidative stress. There's currently ( to my knowledge) no literature to support this theory; however, to me it makes sense.
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