So I got it wrong and I added too little DMSO when I prepared my cell freezing media (100 µL DMSO to 4.9 mL of DMEM, instead of 500 µL DMSO to 4.5 mL DMEM). They are right now in a -80 ºC freezer and I was going to put them in the -140 ºC one, but I'm not sure I should try something or just throw them away.
Is it worth to thaw the vials (2 million cells per vial) and add like 90 µL of DMSO to each vial (to correct the concentration) or is it too late for those little ********?
I would not add DMSO to already frozen cells. DMSO is usually added to prevent cells from water crystal formation that could damage the cells. If you still have growing cells, it should be better to freeze them using 8-10% DMSO, and not use already frozen ones in 2% DMSO. Some people tested the effect of different concentrations of DMSO on cell viability and possible changes: Article Use of different DMSO concentrations for cryopreservation of...
Hi Ignacio, don't throw them away. I would thaw out those cells, have them recover in tissue culture media and then freeze them in 10% DMSO.
Thaw one vial and see if they grow, than decide for the rest of vials,
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