RPMI 1640 Medium (Roswell Park Memorial Institute)

Rpmi-1640 culture medium was originally specifically designed for lymphocyte culture and is now widely used in the culture of a variety of normal and cancer cells, including HeLa cells, Jurkat cells, McF-7 cells, PC12 cells, PBMC cells, astrocytes and cancer cells. It is one of the most widely used media. The unique effect of RPMI RPMI 1640 medium compared to other media is due to the fact that it contains the reducing agent glutathione and a high concentration of vitamins. RPMI 1640 culture medium contains biotin, vitamin B12, and PABA, which are not found in Eagle’s MEM and DMEM.

Ingredients of RPMI 1640 Medium

L-glutamine (Glutamine)

L-glutamine (Glutamine) is an essential nutrient element in cell culture, but it is unstable in solution and will degrade spontaneously. The amount of L-glutamine can be arbitrarily adjusted according to the research needs of the medium without L-glutamine, and the addition of fresh L-glutamine or its substitutes at the time of use is more beneficial to the cell growth.

L-alanyl-L-glutamine (Ala-Glu), also known as alanyl-glutamine and alanyl-glutamyl dipeptide, is an advanced cell culture additive that can directly replace L-glutamine in the cell culture medium. l-Glutamine is an essential nutrient in cell culture, but it is unstable in solution and will, however, it is unstable in solution and spontaneously degrades to produce ammonia and pyroglutamic acid, of which ammonia is harmful to cells, whereas L-alanyl-L-glutamine is very stable in aqueous solution and does not degrade spontaneously. The mechanism of cellular utilization is that during cell culture, cells will gradually release a peptidase into the culture medium to hydrolyze L-alanyl-L-glutamyl into L-alanine and L-glutamine, and then cells will take up and utilize these two hydrolysis products. The process of cellular utilization of L-alanyl-L-glutamyl is similar to the flow-addition culture strategy, in that low levels of L-glutamine are continuously added to the culture medium, thereby increasing the utilization of L-glutamine without generating excess ammonia, which is more conducive to cell growth. L-alanyl-L-glutamyl can replace equimolar L-glutamine, is suitable for all cells, requires little adaptation, and can extend the cell culture time. It can also extend the cell culture time and reduce the number of passages, which saves time and money. Cells cultured in medium supplemented with L-glutamine showed a slower reduction in activity than those cultured in a medium supplemented with L-glutamine. The slightly longer delay period is due to the time required for the release of peptidase and digestion of the dipeptide.

NEAA (Non-Essential Amino Acids)

NEAA (Non-Essential Amino Acids), which are 7 NEAAs of L-alanine, L-glutamic acid, L-asparagine, L-aspartic acid, L-proline, L-serine and glycine added to MEM medium, can reduce the side effects of cells’ own production of non-essential amino acids during cell culture and effectively promote cell proliferation and metabolism.

Glucose can be adjusted at will according to research needs, which is convenient and fast. High sugar type medium is commonly used for fast-growing cells with low adhesion, myeloma cells of hybridoma, clonal cells, transformed cells of DNA transfection, various primary virus host cells, the culture of single cells and vaccine production, such as the use of CHO cells to express EPO and production of hepatitis B vaccine. Low-sugar medium is more suitable for the culture of slower metabolizing, adherent-dependent cells.

HEPES is an excellent biological buffer, which has no toxic effect on cells. The medium with HEPES can maintain a constant pH range for a longer period of time, which can effectively prevent the cell growth from being adversely affected by large pH fluctuations of the culture medium. It can be used in CO2-free incubators.

Phenol red is used as a PH indicator in the culture medium to continuously monitor the pH of the culture medium. Phenol red makes the culture medium yellow at low PH, and purple at higher PH, and red at PH 7.2~7.4, which is most suitable for cell culture. However, phenol red also has some disadvantages. Studies have shown that phenol red can mimic the effects of steroid hormones (especially estrogen), so when using estrogen-sensitive cells (such as breast tissue), it is best to use a medium without phenol red. Phenol red can interfere with detection during flow cytometric analysis. In addition, the presence of phenol red in some serum-free medium formulations can interfere with sodium-potassium balance.

Penicillin-streptomycin Solution

Penicillin-streptomycin mixture, also commonly referred to as “dual antibiotics”, is the most commonly used antibiotic in in vitro culture to prevent microbial contamination, in which penicillin interferes with bacterial cell wall synthesis and is particularly effective against Gram-positive bacteria, while streptomycin inhibits bacterial protein synthesis by binding to the 30S subunit of the bacterial ribosome and is particularly effective against Streptomycin binds to the 30S subunit of the bacterial ribosome and inhibits bacterial protein synthesis, which is effective against both Gram-negative and Gram-positive bacteria, but is particularly effective against Gram-negative bacteria. The combination of penicillin and streptomycin can prevent most of the bacterial contamination.

This product contains amino acids, vitamins, inorganic salts and many other components required for multi-culture of cells, but does not contain proteins, lipids or any growth factors, so the product should be used with serum or serum-free additives.

Cautions

This product has been filtered and de-bacterized and should be used with care for aseptic operation to avoid contamination.
Do not freeze-thaw the product in order to maintain the best use of the product.
This product is intended for scientific research or further study use only, not for diagnosis or treatment.