I tried to thaw my HaCaT cells several times but 99% of them died. They were frozen with 10% DMSO + 10% FBS + 80% DMEM. I also tried 10% DMSO + 90% FBS then they died less but still only 10-20% survived. I centrifuged them at 300g x 5min, RT as suggested by the cell supplier. Other procedures are as standard protocols. Are there any suggestions to keep more cells survive? Thanks a lot!
Ada Tse
Omar Mohammed
Thank you very much for your answer. My cells were 90% confluent and seeded in at least 2 millions/vial and stored in Mr. Frosty with isopropanol at -80oC overnight before going into liquid nitrogen. 300g x 5min is suggested by the cell supplier. I never have problems with other cells before but those are tumor cells. But I can try lower or short centrifugation as you suggested.
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