Can someone please answer me this. I am storing the cells dissolving it in the freezing media(culture media + 5% DMSO) at -80 deg C. Is it safe to do?.
Hi Kanaka Sai Ram Padam ,
Dear Sai Ram,
1. It is not recommended to store the cells in culture medium
2. For freezing your cells, you can use FBS + DMSO (9+1).
3. This combination can be used for storing the cells at deep freezing points.
Thank you,
Good luck
I have previously stored cell lines at -80C. I used 10% DMSO, 70% FBS and 20% media. During culturing the thawed cells, I would avoid centrifugation to remove DMSO. I would wait for 4-5 hours for the cells to adhere and simply change the media. The cell morphology were good. I used to aliquot a lot so that I reduce the freeze thaw cycle as much as possible. Please note that it's not recommended but still can be done if you don't have Access to liquid nitrogen. Hope it helped.
I tested this once: I had several lines like HEK, CHO, 3T3, K562 and a bunch of different cancer lines, mouse and human. All lines declined in numbers within months, at different rates, but this did not seem to affect the characteristics of the surviving cells. Healthy populations could generally be recovered up to 1-3 years, depending on cell line. All vials became sterilized after that.
A cryobiologist friend of mine told me that the mechanism that's killing them is likely slow "ice recrystallization" that happens at -80, but not at -196.
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