Directions
Dissolve 10.0 g of the medium in distilled water and adjust the volume to 1,000 mL. After stirring on a magnetic stirrer for about 30 minutes, sterilize by
autoclaving at 121°C for 15 minutes. Cool the sterilized medium to room temperature and add a roper amount of sterile 10% sodium bicarbonate solution. (pH will be 7.1-7.4 at 37°C in an atmosphere of 5% CO2 in air by adding 12-20 mL of sodium bicarbonate solution.) This mixture should be stopped tightly and stored in a cool, dark place. At the time of use, aseptically add 0.584g L-glutamine (sterilized) and the desired amount of serum.
Summary
Dulbecco’s Modified Eagle Medium “Nissui”② is a powdered medium prepared according to research by R. Dulbecco et al. Up to now, such media were not
autoclavable because of decomposition or deterioration of the constituents, so that sterilization was accomplished by filtration using a membrane filter. Nissui Pharmaceutical Co., Ltd. Succeeded in preparing an autoclavable powdered medium. This medium is specially manufactured so that constituents of the medium and growth of cells with this medium are not impaired. Compared with Eagle’s MEM, this medium contains about twice the concentration of amino acids and about four times the concentration of vitamins. It contains Glycine and L-serine, which are nonessential amino acids; Pyruvate, which is important for glycolysis; and a very small amount of ferric ion. It is nutritious for the growth of cultured cells and is widely used for research on viruses, primary cell cultures, and single
cell cultures.
Storage and Expiration
Close tightly and store at 2-5°C in a dark and dry place.
May be stored up to one (1) year.
Precautions
Do not use the product after its expiry date. Quality of the product is not warranted after its shelf life. When if medium or reagent touched eyes or mouth,
immediately wash with plenty of water, and consult a physician. Any medium, reagent and materials must be sterilized by autoclaving or boiling water after use, and then disposed as industrial waste according to the law or regulations related to dispose.