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Freezing cells in DMSO straight into minus 80?

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4 replies to this topic

#1 science noob

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Posted 18 January 2012 - 07:38 PM

Has anyone stored mammalian cells straight into -80 C without using a gradual freezing system (isopropanol)?

#2 madelingirly



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Posted 19 January 2012 - 01:03 AM

My friends in Egypt, Especially in Egyptian National Cancer Institute freeze cells by putting them in -20 for 5-18 hrs, than directly in Liquid Nitrogen storage, I know it is not as basic freezing protocols but it works fine with them.
I tried and did it here, and when I thawed my cells they were very fine (According to my little experience)

#3 Biouday



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Posted 19 January 2012 - 02:44 AM

Has anyone stored mammalian cells straight into -80 C without using a gradual freezing system (isopropanol)?

No, If anyone does it, they have less chance to recover the cells.

If you donot have Isopropanol gradual freezing system. One can use Thermocol Box. Keep or insert the vial in to thermocol box in a way the atleast the liquid part goes into the thermocol, some time i use to do it when i donot have enough space in gradual freezing system.

Edited by Biouday, 19 January 2012 - 02:45 AM.

#4 rhombus


    Rhombus/Uncle Rhombus

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Posted 19 January 2012 - 02:52 AM

Has anyone stored mammalian cells straight into -80 C without using a gradual freezing system (isopropanol)?

Dear science noob,

You cannot freeze cells straight into a -80 freezer. Generally these days people use a "Mr. Frosty (£50) or a controlled rate freezer machine (£15-20K). I have my own method which costs next to nothing:-

Cells in DMSO/Glycerol into ice for 10 minutes
Then put the cells in a polystyrene box, with a lid. Perforate the box with the tip of a small pair of scissors. Fill the box with cheap tissue paper and put the vials into it......then put the box into a -20 freezer for 2 hours.
After 2 hours put the box into a -80 freezer and leave overnight.
Next day transfer the frozen vials into a liquid nitrogen repository.

Always works for me....usually 90% viability when next initiated into a TC flask.

Hope this is useful

Kindest regards

Uncle Rhombus

#5 bob1


    Thelymitra pulchella

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Posted 19 January 2012 - 12:49 PM

I concur with Rhombus, and have used systems close to the one he describes with similar success. I now use a Mr Frosty, which is also a good cheap alternative. You could probably make a Mr frosty without too much difficulty - all you would need is a plastic sealable container and some way to suspend the cells in the IPA (small tube rack or similar perhaps).

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