Resuscitating a Hybridoma line

[Jorg Kirberg]

In article <D61BoM.930 at dorite.use.com>, crm at iquest.net (pogo) wrote:
> In article <gustilo-2303951334480001 at renal3.med.upenn.edu>, 
> gustilo at pobox.upenn.edu says:

> >I'm trying to resuscitate a hybridoma line that we have. I usually grow
> >the lines in DMEM HG, 10% FCS, HT, Non-Essential AA, Sodium Pyruvate,
> >HEPES. The other lines grow well in this medium but there is one in
> >particular that just won't grow.
> >

> I would try as many things as possible all at once if you are 
> desperate,,,add IL-2, Il-12, GMCSF and up the FBS to 15 or 20%
> You might want to try using IMDM and since some lines prfer a more acidic 
> medium try a small flask at 10% CO2.  Also some cells simply take a long 
> time to recover from freezing--if only a few cells are viable you may need 
> to put the flask in the back of the incubator for about 2 weeks before you 
> see them acively growing.  It sounds silly, but it has saved me several 
> times.  To really help you, I would have to know the parental line--that is 
> your best clue to what is needed.  Good luck from IU research.

Well, trying too many things might not help: When we send hybridoma cells
to other labs, we prefer to send them coming from a growing culture at RT
as frequently cells dont make the change from IMDM to RPMI immediately.
(out of topic: then the recieving lab just adds their medium gradually
over a period of time). The other way from RPMI to IMDM I had problems
once, so rather do not change the medium and keep the one that cells were
growing in before freezing.
In some posting IL-6 was mentioned, and this helped a lot to increase
viability in my hands.

good luck
jorg

-- 
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Joerg Kirberg                     EMAIL: kirberg at bii.ch
Basel Institute for Immunology    FAX:   41-61-605 13 72
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