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
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2 replies to this topic
#2
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Why does a science forum not have pictures of mice and rats?
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.org/wiki/Python_(programming_language)" target="_blank">http://en.wikipedia.org/wiki/Python_(programming_language)</a>, accessed 25June2009.
#3
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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.
