Resuscitating a Hybridoma line

Fri Mar 24 07:41:56 EST 1995

> hi,
> 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.
> The line grew fine in the usual medium and was then frozen in LN2 (3-4
> years ago; before my time). Cell viability (by Trypan Blue exclusion) was
> fine on thawing. Now, it just won't grow in our usual medium.
> Are there any other supplements I should throw in to kick start this
> particular line?
> I have gotten suggestions to add CEGABA, T/B cell growth factors (IL2, 4,
> & 6), conditioned media.... etc.
> Which supplement worked the best for you? I'm running out of vials of this
> line and I can only try a supplement or two.

IL-6 has worked well for me in these situations.  Something else you 
might consider is to inject the hybridoma into pristane-primed nude 
mice.  In vivo conditions may provide multiple factors that cannot be 
duplicated with recombinant cytokines.


Mitch Magee
Department of Research Immunology
Texas Center for Infectious Disease
2303 S.E. Military Dr.
San Antonio, TX  78223-3597

Phone:  (210) 534-8857 xt 209
Fax:    (210) 532-7791
Email:  MMAGEE at tcid.tdh.texas.gov

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