Currently Im dealing with IFN and some cell lines.. I want to keep my cells in -80C and run my experiment tomorrow.
However, I put my cells into the fridge without stimulating my cell with IFN and I was told that I should stimulate the cells with IFN prior storing them into -80C.
So I take out my cells from the fridge (after 5 minutes putting them in the fridge) and stimulate them with IFN.
My question is:
1. What will happens to the cell when I store them in -80C?
2. After stimulating the cell with IFN, are the signal or proteins still living after I store them in -80C?
3. Can I freeze them without adding any cryopreservation medium and stimulate the cells with IFN the next day?
It is highly recommended to add some cryoprotector like DMSO when you freeze your cells, otherwise, you'l have the formation of water crystals that will tear up the membrane of your cells. therefore, all your cells will be dead.
When you store them properly, the freezing is putting quite a lot of stress to your cells, so they won't be the one you want to analyse, you should always give them some calm after thawing, so they can be healthy again.
Is it really required that you freeze them before your experiment?
@Thibault Bouderlique
Thanks for the reply (sorry if I duno how to tag or reply, still new to research gate).
I have 2 cell lines to run my western blot.. however, 1 cell lines reach about 90% confluency at 6x well plate while others reach only 60%.
So I decide to freeze the 90% confluency for 1 day so that I can have both cell lines in similar cell numbers..
My supervisor told me that I should treat the cells with IFN before I freeze them, as the protein or signal wont be dead even the cell dead (without adding any cryoprotector). However I could not find any journal or article to support the explanation.
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