Does slow cooling work better or fast cooling? Is there any specific cooling rate one needs to keep in mind while cryopreserving the U2OS cell line? Also, what concentration of FBS should be optimal for storing U2OS cell lines?
Hello Aditi Gupta
Does slow cooling work better or fast cooling? Is there any specific cooling rate one needs to keep in mind while cryopreserving the U2OS cell line?
Slow rate of freezing at an optimum cooling rate of -1 degree celsius per minute is needed for U2OS cell line. The rate of cooling controls the size of the ice crystals and the rate at which they are formed, both of which may affect cell recovery. Thus, a slow uniform cooling rate of -1°C per minute from ambient is effective.
To achieve this you may store the cryogenic vials in an isopropanol-containing cryo-freezing container such as Mr. Frosty and place them overnight in a -80°C freezer. This will allow the cells to freeze at approximately -1°C/minute, which is ideal for freezing most cell types. You may transfer the cryogenic vials to a liquid nitrogen tank the next day.
What concentration of FBS should be optimal for storing U2OS cell lines?
The freezing medium used for U2OS cell line is complete growth medium supplemented with 5% DMSO. The complete growth medium contains 10% FBS.
Best.
Thank you, Malcolm Nobre for your reply.
I find U2OS to grow slower and poorly compared to HEK293, is it a general phenomenon or am I missing something?
For U2OS is use cryomedium containing 20% FBS and 10% DMSO in DMEM and place tubes in Mr. Frosty container at -80°C which allows a slower reduction in temperature.
U2OS grow slower than HEK293 or HeLa cell lines.
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