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How to remove DMSO after thawing? - (Dec/25/2006 )

DMSO is used as a cryoprotectant in my expreiment, how to i remove it after i thaw the cells? What are the undesirable effects that will affect the cells should it be left with the cells?
thanks

-backlash-

DMSO is cytotoxic in very low concentrations (below 1% if I'm not mistaken), so if you don't remove it, your cells will die.
Way to remove it is to "wash your cells" after thawing. Just put them in a 50 ml tube filled (or a smaller tube) with PBS or medium and spin them down in your centrifuge, discard supernatans and add fresh medium to them and incubate as normal.

-vairus-

For many cell types, both wash or no wash are OK. Thawed cells are fragile and is very sensitive to centrifugation. So I just plate thawed cells in completed medium, when cells are attached, I sometimes change the medium.

-pcrman-

we thaw cells and leave it for 2-3 hrs till the cells stick to the flask and then change media. It works nicely and doesnt seem to affect the cell growth.

-scolix-

QUOTE (vairus @ Dec 26 2006, 10:04 AM)
DMSO is cytotoxic in very low concentrations (below 1% if I'm not mistaken), so if you don't remove it, your cells will die.
Way to remove it is to "wash your cells" after thawing. Just put them in a 50 ml tube filled (or a smaller tube) with PBS or medium and spin them down in your centrifuge, discard supernatans and add fresh medium to them and incubate as normal.


DMSO becomes cytotoxic at increasing concentrations; DMSO becomes critical >0.5 %

-The Bearer-