I am reviving EAHY926 from -80 degrees at passage 38 and I centrifuge to get rid of the DMSO and yet the cells never attach so what could be the problem?
BTW: I am using 10 % DMSO in DMEM while freezing.
Mist likely the vial you got is not the best quality and maybe was not stored correctly and the cells just died. Did you check viability?
Thank you Monika for your answer.
So you are suggesting the cells weren't viable to begin with, so would you mind telling me an easy technique to check cell viability?
Thanks again.
usually due to low amount of isopropanol in your Frosty. causes them to snap freeze instead of gradually cool down which destroys they're ability to reattach and grow
just a thing..ATCC states that for freezing you need to store in complete medium (DMEM+10%serum) plus 5% DMSO , not 10%...high amount of dmso can be toxic. Usually I froze cells (HEK, HeLa, etc...) in 10% DMSO, so it is not so high, but maybe for this cells (that I do not know) could be too much high...
to try to revive ..make an additional wash in complete medium ...and also before centrifuging, dilute them in 10ml of complete medium...
hope could help ...goodluck!
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