Dulbecco's Modified Eagle Medium (DMEM), with 4.5gms Glucose per liter and L-Glutamine, without Sodium bicarbonate and Sodium pyruvate, 20 L

Dulbecco's Modified Eagle Medium is one of the most widely used modification of Eagle's medium. DMEM is a modification of Basal Medium Eagle (BME) that contains four fold concentration of amino acids and vitamins. Additionally, the formulation also includes glycine, serine that contains four fold concentration of amino acids and ferric nitrate. The original formulation contains 1000mgs glucose per litre and was originally used to culture embryonic mouse cells. DMEM high glucose is a further modification of original DMEM and contains 4500mgs glucose per litre. The additional glucose has proved to be useful in cultivating various other cell lines including primary cultures of mouse and chicken cells as well as various normal and transformed cell lines. C985E77, C985E78 and C985E79 is DMEM with 4.5gms glucose per litre and Lglutamine. It does not contain sodium pyruvate. 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 upon nature of supplement added to the medium.