Graham Dellaire popa0206 at PO-Box.McGill.CA
Tue Jul 18 22:48:06 EST 1995

In my lab we freeze cells as follows

trypsinize and spin down 1000 rpm 5 min
take cells up in 1 ml of DMEM with 20% FBS (cold)
add drop wise 1ml of Freezing medium (DMEM 20% DMSO)
resuspend gently and aliquote into two cryotubes (ie one ml each)
*pre chill cryotubes to atleast -4 but -20 or -80 is better

then there are many freezing/cooling chambers available
that utulize isopropyl alcohol etc...

the easiest is to put your cells in a styrofoam tube box (like for PCR products)
and put this at -80 c overnight 
in the morning place in liquid nitrogen

Ideally your cells should drop temperature by 1 degree a minute 
also you may want to change FBS content or resuspend cells in original 
supernatent as it may contain extracellular factors that may help your cells survive the

hope this helps


Graham Dellaire			    Snail Mail:
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