Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

Reviving a hybridoma from LN2

  • Please log in to reply
2 replies to this topic

#1 polaris



  • Active Members
  • Pip
  • 10 posts

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?



#2 lab rat

lab rat

    Why does a science forum not have pictures of mice and rats?

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 245 posts

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



  • Active Members
  • PipPipPipPipPip
  • 31 posts

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.

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.