Hi,
I'm trying to induce cellular senescence in HaCaT cells.
I tried H2O2 and then tried to check the cellular senescence using S A beta gal assay.
Unfortunately I didn't succeed to get SAbeta gal positive cells so far.
Does anyone has a good protocol to induce cellular senescence in HaCaT cells? using H2O2 or other methods?
Thank you in advance.
Hello Imen Jebri
There are a number of stimuli that can induce cellular senescence. You can try using other methods like replicative senescence or senescence induced by ionizing radiation.
Please refer to the research article below for the detailed protocol.
Article Techniques to Induce and Quantify Cellular Senescence
I hope it is helpful.
Good Luck.
Hi Imen Jebri . I'm not familiar with HaCaT cells, but inducing senescence in HUVEC cells is easy. Let them overgrow past confluence. They are contact inhibited, stop proliferating and just sit there.
I do not use HaCaT cells but I used to work on stress resistance on primary dermal cells keratinocytes, fibroblasts, etc. They just undergo senescence when they are cultured after passages. It looks like UB-B is a commonly used inducer for HaCaT. That would give photodamage to DNA. If you want to stick to oxidative stress, UV-C would work but I do not find any study.
Note that HaCaT cells seem to be a cell line that has mutations to be established. That means it would be somewhat different from normal cellular senescence. So I would recommend being careful about the possibility of artifacts in immortalized cell lines.
Thank you for your feedback. Etopside treatment seems to work. I will try UV too.
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