A couple of suggestions:
(1) Increase the serum concentration to 10%. Be sure you are using
high-quality, hybridoma-tested FBS.
(2) After thawing, check viability using trypan blue exclusion.
(3) Consider growing the cells in smaller flasks (T-25 or even multi
well plates. also consider growing the hybridomas over a feeder layer
of resident peritoneal macrophages or mouse fibroblasts. This will
also increase the amount of Ab secreted.
Good luck!
Jay Mone'
