Iscove's Modified Dulbecco's Medium (IMDM), with L-Glutamine and 25mM HEPES Buffer, without Sodium bicarbonate, 1 L

Iscove's Modified Dulbecco's Medium is an enriched modification of Dulbecco's Modified Eagle's Medium wherein serum can be partially or totally replaced by chemically defined substances. The medium contains additional amino acids, sodium selenite, sodium pyruvate, vitamins and inorganic salts. Potassium nitrate is substituted by ferric nitrate. IMDM was the first medium utilizing HEPES buffer. The medium when appropriately supplemented supports good growth of precursor cells of erythrocytes and macrophages. The medium also supports good growth of T and Blymphocytes and a variety of hybrid cells under serum free or reduced serum conditions. C985G02, C985G03 and C985G04 is Iscove's Modified Dulbecco's Medium with L-glutamine and 25mM HEPES buffer. HEPES, a zwitterionic buffer having a pKa of 7.3 at 37ºC prevents the initial rise in pH that tends to occur at the initiation of a culture and increases the buffering capacity of the medium. 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.