cryopreserving hybridomas

Stephen Gisselbrecht gisselbr at husc8.harvard.edu
Tue Aug 24 14:43:13 EST 1993


In article <9308241751.AA10294 at post.its.mcw.edu> fgarbrec at FRED.BMT.MCW.EDU writes:
>I have, over the past couple of years, gotten a
>number of murine hybridomas to grow well in various
>serum free media, but have been unsuccessful in 
>cryopreserving serum-free adapted hybrid cell lines.
>I use the standard 10%DMSO/90%FCS as a cryopreservative,

	I can't tell you what to do from experience, since I have none,
but I'll bet you money (and I'm a grad. student!) that you could get
around this by slowly readapting your cells to FCS.  Maybe make a standard
RPMI+10%FCS, passage them into 90%SFM/10%RPMI+FCS, next day go to 80/20,
etc.  Within a week and a half you should be able to freeze them in
FCS+DMSO, and then reverse the procedure when you thaw them.
	Not as pleasing an answer as some formulation that would allow you
to freeze your cells as is, but I don't have one of those, and I thought
an inferior suggestion would be better than none at all.

					steve gisselbrecht
					cell & dev. bio.
					harvard medical school



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