Hybridoma development - (Jun/30/2012 )

Hi all,
Very recently I am observing that fusion plates are forming clumps ( 2nd/3rd day) and hybridomas started dieng.... I tried changing all possible reagents and even new vial of Sp2/O has been thawed freshly for fusion though the result is same, cell started clumping, but the control wells are fine (where only spleen and only Sp2/O plated in HAT). Spleen is from immunized Balb/C mice... Can any one please suggest .....

Hi, probably it's too late anyway...did you check mycoplasma contamination? This is one of the most common reasons of no hybridoma development