Why use DMSO and FCS for freezing cells? - (Feb/26/2005 )
Hi!, anyone know the theory besides why we use DMSO and FCS for freezing??
I do not know the exact role of FCS but DMSO is used as a cryoprotectant. it prevents formation of ice crystals which otherwise lyses the cells during thawing. May be FCS is used as a stabilizer since it contains abundant proteins.
use of FCS is not absolute necessary, due to the fact m boss use serum free medium + DMSO in order to freeze his cells in nitrogen....
seems that using DMSO is more important.
moreover in the atcc site, there are guidelines for freezing cells
using dmso is as they explained before, using fcs is importent too for 2 reasons one of them is for dilluting the dmso (dmso can hurt the envelope of the cells thats why u freez the cell with 10% dmso and the rest is fcs or fbs- it doesnt matter)- so when u thaw the cells they will be ok
I agree with Fred in that FBS or FCS are not necessary. I froze the cells in 7.5% DMSO diluted in DMEM and they are OK at thawning. Maybe for particular cell lines is better to use FBS/FCS, but for most cases is just a waste of money, as DMEM works equally well.
DMSO as has been stated in several reserach papers is able to prevent crystalization of water that will cause cells to lyse during crypreservation. This is usally a 10% solution in a medium containing up to 20% of a serum (I use horse serum because it is inexpensive compared to calf/bovine serums). The reason for the serum is so when thawing the cells they are not going to be starved of nutrients and it also acts as a cushion for the cells during the thawing process depending on the amount of time it takes to take the thawed vial of cells to reach a culturing plate in fresh medium. Because DMSO is toxic to the cells in an extended amount of exposure it is important to centrifuge the cells into a pellet and remove as much DMSO containing medium as possible and replace it with fresh growth medium when plating.