Nutrient Mixture F-12 Ham, with L-Glutamine, without Sodium (C08-0442-754)

Ham's Nutrient Mixtures were originally developed for single cell plating of near diploid Chinese hamster ovary (CHO) cells and mouse L-cells. Both F-10 and F-12 are formulated for use with or without serum, depending on the type of cells being cultured. Ham's Nutrient Mixture F12 was originally designed for serial propagation and cloning of two CHO cell lines namely, CHD-3 and CHL-1 and mouse L cells. It is the medium of choice for the growth of cells of rodent origin and for cloning of myeloma and hybridoma cells. This medium is also the medium of choice for clonal toxicity assay using CHO cells. C985F03, C985F04, C985F05 and C985F06 is Nutrient Mixture F-12 Ham with Lglutamine. Users are advised to review the literature for recommendations regarding medium supplementation and physiological growth requirements specific for different cell lines. Storage and Shelf Life 1. All the powdered media and prepared liquid culture media should be stored at 2-8°C. Use before the expiry date. Inspite of above recommended storage condition, certain powdered medium may show some signs of deterioration /degradation in certain instances. This can be indicated by change in colour, change in appearance and presence of particulate matter and haziness after dissolution. 2. Preparation of concentrated medium is not recommended since free base amino acids and salt complexes having low solubility may precipitate in concentrated medium. 3. pH and sodium bicarbonate concentration of the prepared medium are critical factors affecting cell growth. This is also influenced by amount of medium and volume of culture vessel used (surface to volume ratio). For example, in large bottles, such as Roux bottles pH tends to rise perceptibly as significant volume of carbon dioxide is released. Therefore, optimal conditions of pH, sodium bicarbonate concentration, surface to volume ratio must be determined for each cell type. We recommend stringent monitoring of pH. If needed, pH can be adjusted by using sterile 1N HCl or 1N NaOH or by bubbling in carbon dioxide. 4. If required, supplements can be added to the medium prior to or after filter sterilization observing sterility precautions. Shelf life of the medium will depend on the nature of supplement added to the medium.