Two-step freezing of hybridoma cells in 96-well microculture plates
Language English Country Czech Republic Media print
Document type Journal Article
PubMed
4065377
Knihovny.cz E-resources
- MeSH
- Fibroblasts cytology MeSH
- Cell Fusion MeSH
- Hybridomas cytology MeSH
- Lymphocytes cytology immunology MeSH
- Mice MeSH
- Plasmacytoma immunology pathology MeSH
- Tissue Preservation instrumentation methods MeSH
- Freezing MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
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.