The effect of postfusion cell density on establishment of hybridomas

The effect of postfusion cell density on establishment of hybridomas in primary microculture was studied. Following fusion of BALB/c spleen and NS0 myeloma cells in the ratio of 5:1 we demonstrated that seeding the cells at a concentration of about 20-25 X 10(4) spleen cells per well resulted in maximal number of wells containing only one growing hybridoma clone (40.1-43.7% plated wells). Importance of generation of microculture with one clone for the effectiveness of preparation of mAb-secreting hybridomas is discussed.