Biological medium

Most media is based on a balanced salt solution (BSS), which is supplemented with amino acids, vitamins, and other nutrients similar to those in serum. The most widely used medium is a mixture of Eearle`s MEM, which contains 13 essential amino acids and 8 vitamins. The Ham`s F12 also includes non-essential amino acids, and the range of vitamins is also very wide. In addition, the conventional two media containing inorganic salts and metabolic additives MEM / F12 each take 1/2, forming the most versatile medium for neurobiology.

Dulbecco`s modified medium, DMEM, is now applied to rapidly growing cells. It contains the same nutrients as MEM, but the concentration is 2 to 4 times higher. Choose a certain medium, you should carefully understand the composition table, you should know that in most cases, the medium is insufficient. For example, some culture media include glutamic acid in the amino acid, and although this culture medium is widely used in the field of neurobiology, it is sensitive to glutamate-sensitive neurons that may have extracellular toxic damage , Is not the best choice, especially if neurons grow in a lack of gelatinous environment. F12 contains ferrous sulfate and is reported to have neurotoxic effects. In all these media, glutamic acid has a higher concentration than other amino acids because of its instability and its use as a carbon source in many cell cultures. For the cultivation of neurons, the glucose content in the basal medium is often increased to 0.6% or pyruvate is added (if these two substances are lacking in the medium).

Both MEM and F12 should be balanced with 5% CO2. DMEM contains a higher concentration of NaCO3. It needs to be balanced with 10% CO2. Of course, it can also be used at a lower CO2 concentration. The composition of these basic media is based on the study of the growth of different cell lines, but usually used in primary culture can also have relatively satisfactory results. In principle, HEPES can be used as a buffer to replace bicarbonate in order to relieve the limitation of the high concentration CO2 culture environment. It is not so simple in actual operation. Obviously, dissolved CO2 and bicarbonate are important for good cell growth. Leiboviz`s L15 medium can be used to grow nerve cells in an atmospheric environment. The medium uses a distinctive BSS as the basis. It contains high concentrations of amino acids to improve buffering capacity. The medium uses galactose as a carbon source. In order to prevent the formation of lactic acid in the medium, a small amount of dissolved CO2 is produced by pyruvate metabolism. The advantages of this medium are obvious, especially when it is difficult to maintain high CO2, such as in long-term micromanipulation and physiological studies. L15 medium has been used to successfully culture peripheral neurons, but it has not been fully tested in CNS neuronal development studies. Selection of cell culture media selection There is no certain standard for the selection of culture media. There are several suggestions for reference: (1) The culture medium used to establish a certain cell strain should be the first choice for the cultivation of such cells. You can consult the references or consult when purchasing cell lines. (2) Other media commonly used in laboratories may wish to try, many media can be suitable for a variety of cells. (3) Select the medium according to the characteristics of the cell line and the needs of the experiment. Such as mouse cell line, multiple selection RPMI1640

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