Good afternoon everyone,
We're measuring senescence with flow cytometry (with C12FDG), we compare irradiated cells (2 Gy and 5 Gy) and non-irradiated cells (= Control). But we don't see any C12FDG positive cells, not even in the cells treated with 5 Gy. So I'm wondering If maybe our C12FDG is to old. Is there a way to induce senescence in cultured cells, so that we have a positive control for our experiments? Thank you very much :)
Hello Giulia Pfeil
You may use hydrogen peroxide which is widely used to achieve oxidative stress-induced premature senescence within a short period of time. You may start with a dose of 20μM H2O2. For more information you may refer to the article attached below. Please see Fig 6 which shows detection of SA-β-Gal in the MGFs cells using 20 μM H2O2.
Article Oxidative stress caused by a low concentration of hydrogen p...
Best.
@Malcom Nobre: Thank you very much! :) I'll try it next time. Have a nice weekend :)
Hi Giulia,
In our lab we have established a very reliable in vitro model to induce permanent SIPS using Doxorubicin. Several markers of senescence were upregulated.
... H2O2 was only semi-successful in our hands. Some, but not all senescence marker could be confirmed. This might depend on cell type, concentration of H2O2 and exposure time.
Please find attached the publication on the doxorubicine-based model.
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