I have been growing my hybridoma cells inGibco Hybridoma SFM . I have cryopresrved them using 92.5% medium (50:50 ratio of fresh to conditioned media) +7.5% DMSO , the viability was 87%.
i froze them gradually using -80 overnight then in the liquid nitrogen.
but when i revived them they died. I revived them by rapid thawing at 37 C then by putting them in 20 ml pre-warmed fresh SFM.
I also tired reviving them using serum supplemented Media but they also died.
I tried by both to centrifuge the thawed cells and not centrifuging them .
Can any one help me with the protocol to revive them .
Or should i have used a different cryopreservation protocol.
Hi, Hadeel.
I think you have to find optimum cell freezing and thawing. Most of cells are damaged during freezing and thawing. Refer a basic mammalian cell culture book. Read it through the freezing and thawing cell sections. Once you get familiarized all the precautions, it's nothing.
Best wishes.
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