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cryopreservation agent missing in frozen cell stock - (Nov/28/2006 )

i am very new to cell culture and neglected to add a cryopreservation agent (DMSO or glycerol) to cells before freezing them. my freezing procedure was simply to aliquot cells (1.5 ml) and allow to freeze at -80C for 2 days, then move to LN storage. i just thawed cells yesterday to start culturing and they look terrible; are cells likely to recover at all without having been preserved properly when frozen and stored?


well the answer is strongly dependant on cell type
for primary cell lines i think it's no.
For strong tumor cells 293 or HeLa, chances are 40% max
for the other cell types i don't really now.
I think endothelial cells are very fragile as well.

see that link for tips about cryopreservation.
also available in french


Most likely these will not make it. Without DMSO, ice crystals can rupture cell membrane easily. Few will survive.


even absence of a cryopreservant affect the homeostasis in the cells due to water absence..