Two-step freezing of cells used in hybridoma technology.
Hybridoma technology requires freezing of parental myeloma cells, continuous freezing and thawing of clones and stable hybridoma cells. A modification of the method of two-step freezing is described. The cryoprotective agent (5% dimethyl sulphoxide) is added to the cells at room temperature for 10 min. Cells are then transferred directly to the -25 degrees C bath, held at this temperature for 10 min, and stored directly in liquid nitrogen. Thawing is rapid in a water bath warmed to 60-80 degrees C. Hybridoma cells retain high viability and the production of specific monoclonal antibody after thawing.
D000818:Animals; D002470:Cell Survival; D005347:Fibroblasts / Q000166:cytology; D005615:Freezing; D006822:Hybrid Cells / Q000166:cytology; D006825:Hybridomas / Q000166:cytology*; D051379:Mice; D010954:Plasmacytoma / Q000473:pathology; D013154:Spleen / Q000166:cytology; D014021:Tissue Preservation / Q000379:methods