Hybridomae

Kiley R. Prilliman Kiley-Prilliman at ouhsc.edu
Wed Apr 15 21:47:44 EST 1998


: > I am getting extremely variable rates of viability from my B cell
lines
: > when I thaw them after storage in liquid N2. I freeze them in 50%
media/
: > 30% FCS/20% DMSO. I freeze them gradually (I use on of those little
: > ethanol-jacked freezing things that claims to give 1oC/hour) and I
thaw
: > very rapidly (37 water bath) and rinse them thoroughly.
: >
: > AND YET--I still get some vials with less than 1% viability while
others
: > are fine. Some lines are better than others, but I see this problem
even
: > within the same line, frozen at the same time. I'm worried about
: > completely losing some of my more fragile lines.

...I typically freeze hybridomas in 40% media/50%FCS/10% DMSO.  I don't
know if your increased amount of DMSO could be causing a problem or not
(probably *not*, there are probably about as many freezing solutions
available as there are people freezing hybridomas!), but some of the
hybridoma lines I have worked with have shown a distinct distaste for
DMSO concentrations above 10%.

Kiley




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