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Reviving a hybridoma from LN2


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#1 polaris

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Posted 20 May 2010 - 07:27 AM

Hi all,

We're about to revive some frozen hybridoma cells and the procedure recommends co-culturing with normal mouse spleen cells until the hybridoma cells can proliferate on their own. I'm having trouble finding a source of normal mouse spleen cells.

1. Is it necessary to co-culture?

2. Sources of normal mouse spleen cells?

3. Alternatively I think we can just use conditioned medium but the source I found (Sigma) has it back ordered until July. Any other conditioned medium sources?

Thanks!

-P

#2 lab rat

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Posted 20 May 2010 - 09:57 AM

We just added some IL-6 to some of our lines to kick-start them. I don't recall co-culturing.
42..."An immutable fixed-precision number of unlimited magnitude." <a href="http://en.wikipedia....amming_language)" target="_blank">http://en.wikipedia....amming_language)</a>, accessed 25June2009.

#3 Roo

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Posted 30 August 2010 - 06:48 AM

Sometimes we co-culture with normal mouse spleen cells, and sometimes we don't. We do, however, make them ourselves. Just sacrifice a "normal" (non-stimulated for Ab production) mouse, preferably the same strain, and dissect out the spleen. Collect/extract the spleen cells the same way you would for a hybridoma fusion (including some washes). Resuspend in medium and plate out. Typically, we do this the day before we need them.




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