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Storing frozen cells in -80C instead of liquid nitrogen

cryopreservation liquid nitrogen -80 degrees cell culture

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7 replies to this topic

#1 distalless

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Posted 20 September 2011 - 03:03 PM

Hi,

I usually freeze my cells in -80 degrees and then transfer them to a liquid nitrogen tank a day later for future storage.

However, I left one of my cell lines in the -80 degrees accidentally for a month so I was just wondering if they will be OK. How do I know if they are not OK?

Thanks a lot!

#2 bob1

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Posted 20 September 2011 - 05:21 PM

Most (non-primary) cell lines will be OK for a month at -80, primary cell lines might have some issues though. The only real way to check is to thaw them out and see if they still grow OK.

#3 distalless

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Posted 20 September 2011 - 06:37 PM

It is a leukemic cell line (HL60 cells). The cells seem to be growing fine, but I am worried that maybe there will be some genotypic changes (my research is in genomics so I would be concerned about sensitive changes) Thanks!

#4 bob1

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Posted 21 September 2011 - 05:33 PM

In that case, no-one will have looked and the only way to tell will be to compare the expression of many genes (perhaps microarray) between cells you know have been treated OK and these ones.

Lab based evolution of cell lines is a big problem...

#5 allynspear

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Posted 28 September 2011 - 07:51 AM

I would say you have a worse chance of generating genomic changes by continuous passaging than you do by one month storage at -80 degrees.

#6 Neanderthal

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Posted 15 November 2011 - 10:06 PM

Some of the people in my department use -80 to store their cells. I have not heard any serious problems with them. But, since your work depends a lot on genotypic stability, unless, as bob 1 said, you do some real experiments to address this problem, I don't think one will be able to tell the genomic changes occurred.

The ideal thing to do is to keep the cells in -80 for a day or two soon after freezing down protocol (in a Nalgene cryofreezer or something of that kind) and then transfer them into liquid nitrogen

#7 h9494036

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Posted 10 September 2013 - 12:22 PM

It is only good for short-term storage.  I believe 6 months are the shelf-life, and longer than that is at your own risk.



#8 everyday lab rat

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Posted 11 September 2013 - 02:58 PM

We unfortunately had to do 'the great -80C experiment' one year.  A -140C freezer went down and the only space available was in a -80C freezer. The cells where mostly human PBMCs and lymphs. After a few months at -80C, the viability of the cells when thawed was miserable. Usually  20-30%V. 







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