It seems also to depends on the cell line. HeLa, HEK293, or U2OS survive longer at -80 than immortalized B cells from what I have observed. Even afte ra year and half at -80C, some of my U2OS cell lines were still alive but with a good 80/90% death rate on thawing so it is likely they had mutations/epigenetic changes (but depending of the experiment you want to perform it can be not important).
-- Freezing medium also play a role in thawing survival. Some of my cell lines survived better in 50% fresh medium, 40% FBS, 10% DMSO than 90% FBS and 10% DMSO for example. DMSO is also toxic for the cells at 37 degrees (should also be at room temperature) which is why the cells need to be put in a medium as soon as possible after thawing (warm medium also improve cell survival). For the freezing medium, it is better to follow what is written on the ATCC website since it is tailored for each cell line.
But yeah as a rule, it is better to keep cell lines in liquid nitrogen.
On Monday, April 20, 2015 at 6:01:47 PM UTC+1, Mega [Andreas Stuermer] wrote:
On Monday, April 20, 2015 at 6:01:47 PM UTC+1, Mega [Andreas Stuermer] wrote:
Hi everyone,
Today at University I heard that human cells from the -80°C freezer often don't work after thawing. Especially when being frozen for a year or more.
I always thought -80°C would work well and -130°C would be better but optional.
Is this due to our freezer, or do you have similar experience?
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