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Two-step freezing of cells used in hybridoma technology

. 1985 ; 31 (5) : 340-7.

Language English Country Czech Republic Media print

Document type Journal Article

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.

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