Medium 199, with Hank's salts and L-Glutamine, without Sodiu (C08-0442-013)

Medium 199 was the first nutritionally defined medium developed by Morgan, Morton, and Parker in 1950. This complex medium was formulated specifically for nutritional studies on primary chick embryo fibroblasts in the absence of any additives. It was observed that explanted tissue could survive in Medium 199 without serum but long term cultivation of cells required supplementation of the medium with serum. Medium 199 is formulated with either Hank's salts or Earle's salts. The medium when supplemented with serum can be used for growth of a wide variety of cells. Medium 199 is presently used for the maintenance of non-transformed cells, vaccine and virus production and primary explants of epithelial cells. C985F84, C985F85 and C985F86 is Medium 199 with Hank's salts and L-glutamine. 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.