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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?

-klb206-

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

-fred_33-

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

-genehunter-1-

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

-strawberry-