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Freezing cells


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#1 doctormoon

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Posted 15 September 2010 - 08:29 PM

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?

#2 Sachin Patil

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Posted 16 September 2010 - 12:07 AM

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?


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

#3 MJD

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Posted 16 September 2010 - 04:59 AM

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.

#4 Rupam

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Posted 16 September 2010 - 05:45 PM

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.

You can also widen your knowledge and clear your doubts by going through test of wiziq through [url="[url]http://www.wiziq.com/tests/biology"]biology[/url] test papers, biology test[/url].It will surely help you.

Regards
Rupam

#5 Hugobox

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Posted 08 October 2010 - 02:51 AM

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.

You can also widen your knowledge and clear your doubts by going through test of wiziq through http://www.wiziq.com/tests/biology.It will surely help you.

Regards
Rupam


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.

#6 LabLackey

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Posted 08 October 2010 - 07:07 AM

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

#7 bob1

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Posted 08 October 2010 - 02:26 PM

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.




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