I am working with raw cell line from last one year and they were fine before .But since last month they keep on self activating.. ( without treating any activating agent like LPS) even after thawing during 1st passage their morphology changes completely from round to star shape.
I have tried using different stocks...
All other cell lines in our incubator are fine so I think its not incubator problem, even then i tried to clean it all over again but all in vain..
what should I try .. ??
did you replace the media as well? FCS for example can have different batch to batch effects. Do you use any plastics which could be contaminated? Test your cells for mycoplasma, its possible that the batch you froze last year had something inside...
One possibility is that might be your media has LPS contamination or if you are using water to dissolve your media (powder), so there are bright chance that water is LPS positive. Incubate your cells with Polymyxin B and then check that cells are activating or not if they are not activating this means your media has LPS contamination.
Thanks guys for your suggestions..
Yes I have tried to make new media but it didn't showed any difference.. I use cell culture flasks which are sterile.. OK i will check for mycoplasma .But all stocks are from different lab members which they made in different time...
We do not use powder media so this can't be the reason..
Do other cell lines in incubator could be any kinnda influence on this ?
If some cells are infected with lets say mycoplasma yes, but i have never heard of cells activating other cells across the incubator.
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