Two-step freezing of cells used in hybridoma technology
Language English Country Czech Republic Media print
Document type Journal Article
PubMed
4065374
Knihovny.cz E-resources
- MeSH
- Fibroblasts cytology MeSH
- Hybrid Cells cytology MeSH
- Hybridomas cytology MeSH
- Mice MeSH
- Plasmacytoma pathology MeSH
- Spleen cytology MeSH
- Tissue Preservation methods MeSH
- Cell Survival * MeSH
- Freezing MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
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.