Resuscitating a Hybridoma line

gustilo at gustilo at
Thu Mar 23 13:34:48 EST 1995


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.

thanks very much in advance.


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    I am not responsible for comments that I make after 3:00 A.M. or after 5 cups of double espresso, whichever comes first.

gustilo at

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