Resuscitating a Hybridoma line

[ANDREI POPOV]

>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.

I would use RPMI1640 (maybe your DMEM is OK)
+ 20% FCS, L-Gln, 2-ME (essential for all lymphoid cell lines)

Find out what was the parental myeloma.
In my hands NS0 did not feel happy in the presence of
Na Pyruvate, though SP2/0 liked it very much.

Try  feeder cells (normally isogenic splenocytes work great)

If all this fails, I would simply inject freshly
thawed cells i/p into pristanized mouse
(hope you are not in UK)


Good luck

andrei
andrei.popov at bbsrc.ac.uk