Has anyone worked on differentiation and H2O2 treatment in H9C2 cells

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Iro Damopoulou
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Has anyone worked on differentiation and H2O2 treatment in H9C2 cells

I differentiate H9C2 cells from myoblasts to myocytes for 7 days in a 96-well plate (10.000 cells/100 μL). On the 7th day i do the treatment with the hydrogen peroxide (500uM) for 12 hours and then i'm checking cell viability with MTT assay. The differences between my CTL group and the treated one suggests that the treatment doesn't work, but when i repeat the treatment in undifferentiated cells in the 96-well plate the treatment works successfully. Any idea about what could be the problem? Maybe the number of the cells i use for the differentiation, the fact that i do the differentiation in the 96-well plate or anything else?Any help would be completely appreciated,thanks in advance