Storing cell lines long-term at -70C
Posted 07 July 2010 - 02:12 PM
A quick question - has anyone had any experience at storing cell lines at -70C as opposed to -140C ..?
I've usually stored such at -140C, but it looks like our access to a -140C freezer is going to be limited for a few months ... and so I was wondering if cell lines could be stored at only -70C, even if only for a limited time. I've done a little interweb snooping on the subject, and have come across very little - i.e. none - mention of storing cells at -70C. Does this mean it's simply not possible ..?
Comments/opinion etc. welcome!
Posted 07 July 2010 - 06:36 PM
Posted 07 July 2010 - 06:46 PM
If I may ask further, has this been the case with a broad range of cell types, or generally with more 'hardy' cells lines versus stable transfection clones, etc?
I'm primarily interested in DU145 and PC3 cells, neither of which I have had much experience in maintaining, and so I'm not sure how they'll fare under sub-optimal conditions.
Again, thanks for any feedback on the matter!
Posted 06 August 2010 - 11:35 PM
I observed some primary cells (HUVEC), that die even after 4 days at -80 °C (viability at thawing <10 %). Transfer the same lot of cells to N2 on the day after freezing leads to a viability >90 %.
In generally you can say, that primary cells are more sensitiv to -80 °C storage than cell lines. But even cell lines might have problems with long-term storage in 80 °C (e.g. Hek293 cells are fine after 1-2 weeks of -80 storage, but after 4 weeks the viability decreases to about 50 %, 8-12 weeks at -80 the viability of the same freezing batch is below 20 %).
Some cells like keratinocytes (primary and cell lines) are highly tolerable to -80 °C storage and nearly don't suffer even of one year storage at -80°C.
I don't have experiance with your cell lines, but just to make shure not to run in problems, try to avoid storage of cells at -80 °C longer than one week. I think that most cell lines even tolerate up to 1 month storage at -80 °C.
Posted 25 August 2010 - 09:46 AM