Two-step freezing of hybridoma cells in 96-well microculture plates.
Stabile hybridoma cells, colonies of hybridoma cells 14 days after fusion of immune spleen and myeloma cells, myeloma cells and fibroblasts cultured in 96-well microculture plates were frozen by the method of two-step freezing. The culture medium was aspirated, and 50 microliter of the medium containing a cryoprotectant (5% dimethyl sulphoxide) was added for 10 min at room temperature. The plates were put into microtene bags, placed at -25 degrees C in a freezer for 30 min and then stored at -100 degrees C in liquid nitrogen vapour. Plates with cells were thawed rapidly in a 50 degree C water bath. After thawing the hybrid cells were viable and continued to produce the specific antibody.
D000818:Animals; D002459:Cell Fusion; D005347:Fibroblasts / Q000166:cytology; D005615:Freezing; D006825:Hybridomas / Q000166:cytology*; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D010954:Plasmacytoma / Q000276:immunology / Q000473:pathology; D014021:Tissue Preservation / Q000295:instrumentation / Q000379:methods