Harold Drabkin: DMSO for tissue culture u

Simon Dawson mbxspd at unicorn.nott.ac.uk
Thu Jun 8 02:52:50 EST 1995



As far as the effects of polystyrene dissolved in DMSO is concerned, I 
guess that it remains in solution and is massively diluted out once you 
add culture medium.
DMSO is however VERY toxic to cells ! I add DMSO which is at room 
temperature, give it 1 brief aspiration with the same pippette, and 
IMMEDIATELY place it on ice and then into a -70 freezer a.s.a.p.
I'm told that you should aim to cool the cells by 1 degree per minute to 
prevent ice formation, tho' in my hands Sigma Mol. Biol. grade DMSO 
placed in a polystyrene rack inside a plastic box freezes the cells 
well.
When it comes to thawing, this is done as quickly as poss at 37 degrees.
I then add growth medium (as much as poss, mix gently and spin out the 
cells. I then resuspend the cells in the working volume of fresh growth 
medium and plate/flask them.
It may be that you have MEGA sensitive cells, but this works for all the 
cell types used here.
Regards,
Simon





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