I am working with ARPE19 cells and have been conducting mito stress tests using seahorse technology to determine the optimum H2O2 concentration for inducing cellular damage, however, my basal respiration for the ctrl is lower than that of the treated cells. 96 well plate seeded with 5*10^4 cells/well , with treated cells exposed to H2O2 for an hour. Could it just be that I'm not plating evenly as the ctrls are plated last? Any suggestions as to what the issue may be?
I wonder if the cells might be experiencing mitohormesis, where the mitochondria are effectively hyperactive in order to counteract a low level/brief stress. You could perhaps try the same dose levels but for a longer time period and look at multiple time points? Hope this helps
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