Freezing cells - How long can I keep cells at -80C? (Sep/15/2010 )

I am freezing cells (A549). I put cells in 10%DMSO-containing medium and put it cryocontainer at -80C.
I am going to relocate it into the -270C nitrogen tank.

My question is how long I can keep cells at -80C before going to -270C. I know it would be 1~2 days.
How about 3 days or 4 days as in my case?

doctormoon on Thu Sep 16 04:29:00 2010 said:

Hi
I haven't done experiments on your cell line but in general the cells can be stored upto 5-7 days at a constant -80 degree. I have done this with Caco-2, MCF. 3T3 without any problem (take care, there should be no temperature fluctuations as someone may open it frequently or may keep open for a long time). Due to this sudden temperature changes cells may get a shock and die.
I will also suggest using DMSO at 5% concentration as DMSO is toxic at higher concentration and is sufficient for storage of cells.
Sachin

I've worked with prostate (DU145, PC-3, LnCaps) and breast cancer (MCF-7, MB-231) cells and these tolerated up to several months in the -80C freezer prior to transfer to the nitrogen tank. When taken out of the nitrogen tank almost a year later, the cells performed the same as they did previously in all experiments. Therefore, you should be fine storing your cells in the -80C for 3-4 days.

You can store the cell for 3-4 days in -80C but the sudden increase in temprature may be harmful for the cell as it cause the cell to die so you have to be little attentive while doing that.

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Regards
Rupam

Rupam on Fri Sep 17 01:45:19 2010 said:

It seems that if the cells store over 3-4 days may cause them to die. I normally back up my cells (Caco-2, Hela, and A549) in -80C freezer more than one month due to lack of nitrogen facilities. I once tried to regrew the Caco-2 cells and it took about 2 month to recover them.

My lab keeps our cell lines (BSR-T7/5, Vero) in the -80. I don't even think we have any in our -140.

Many cell lines will survive up to 6 months (or possibly longer) in the -80, but it is still not a good idea to store them in there for that long as you are then applying a selective pressure on the cells for those cells that can survive the conditions in the tubes (DMSO, crystal formation etc.). There is still some metabolic activity at -80, which is why phenol red truns yellowish with storage in the -80.