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Cell Culture Media

Cell culture media used in laboratory

1. What is cell culture media?
    1.1 Composition and role of cell culture media
2. Type of cell culture media
    2.1 Cell medium
3. How to buy cell culture media?

What is cell culture media?

Cell culture medium is both the basic material for supplying cell nutrition and promoting cell reproduction and proliferation in cultured cells, as well as the living environment for the growth and reproduction of cultured cells.

Composition and role of cell culture media

A. Amino acids

Amino acids are the basic units of protein. Different types of cells have different requirements for amino acids, but there are several amino acids that cells cannot synthesize on their own and must rely on the culture medium to provide them, which are called essential amino acids. Among them, glutamine is an essential amino acid for the synthesis of nucleic acids and proteins, and in the absence of glutamine, cells grow poorly and die. The essential amino acids include L-glutamine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-threonine, L-tryptophan, and L-valine.

B. Vitamins

Biologically active substances that maintain cell growth and play a regulatory and controlling role in cell metabolism. In cell culture, although serum is an important source of vitamins, many culture media are supplemented with a variety of vitamins to suit the growth of a wider range of cell lines.
Fat-soluble vitamins such as A, D, E, and K.

Water-soluble vitamins such as B1, B2, B6, B12, pantothenic acid, folic acid, biotin, C, niacinamide, etc.

Many vitamins are involved in the composition of the active groups of various enzymes, without which the enzymes would be inactive and metabolic activity would not take place.

VA is a carrier of oligosaccharide groups in the synthesis of glycoproteins in cells and has an important role in the maintenance of epithelial cells. vD is involved in the regulation of calcium absorption. ve is an antioxidant that prevents the oxidation of unsaturated fatty acids in phospholipids that make up biological membranes. vK deficiency causes low prothrombin and prolonged clotting time.
Folic acid is an important raw material for the synthesis of tetrahydrofolate, which plays an important role in the biosynthesis of nucleic acids and the biosynthesis of proteins.

Biotin is a component of a number of specific carboxylases involved in the process of sugar metabolism and fatty acid synthesis.

C. Carbohydrates

Carbohydrates are the main source of energy for cell growth, some of which are components of the synthesis of proteins and nucleic acids. The main ones are glucose, ribose, deoxyribose, sodium pyruvate, and acetate.

D. Inorganic ions

Basic inorganic ions such as sodium, potassium, magnesium, calcium, and phosphorus, are necessary for cell composition and are involved in cell metabolism.

The main function of inorganic salts in the culture medium is to help cells maintain osmotic pressure homeostasis. In addition, by providing sodium, potassium, and calcium ions, it helps cells to regulate cell membrane function. The osmolarity of the cell culture medium is a very important factor, and cells can usually tolerate 260 mOsm/kg - 320 mOsm/kg. The osmolarity of the standard culture medium fluctuates within this range. Special Note: The addition of other substances to the culture medium may significantly alter the osmolarity of the culture medium, especially those dissolved in strong acids or bases.

Na+ is the most important cation in the extracellular fluid and plays a decisive role in maintaining a constant osmotic pressure. k+ is mainly distributed in the intracellular fluid, and intracellular k+ is necessary for activating certain enzymes, and it is also extremely important in regulating the acid-base balance of the intracellular environment. ca2+ is used in the extracellular fluid to adhere cells within tissues to each other, and is involved in many important cellular physiological activities within the cell. Mg2+ is an important component of the intercellular matrix and is important for the stable binding of cells to each other. The functions of phosphorus compounds for the regulation of cellular material metabolism and physiological functions are very extensive and indispensable.

Type of cell culture media


Cell medium

There are many kinds of classical culture media, among which DMEM, RPMI 1640, MEM, and DMEM/F12 are the most widely used media. Others such as M199, IMDM, and L15 medium are also used for the culture of certain cells. Their specific characteristics and applications are as follows.

A. BME cell culture medium

Basic Eagle medium, designed by Eagle in 1955, BSS + 12 amino acids + glutamine + 8 vitamins. It is simple, easy to add, and suitable for various passaged cell lines and for special research. The varieties of cell culture mediums improved on this basis are MEM, DMEM, IMDM, etc.

B. MEM cell culture medium

MEM cell culture medium, also known as low-limit Eagle's medium (Minimal Essential Medium), was modified from Eagle's basic medium (BME) in 1959. It has deleted lysine and biotin, the increased amino acid concentration is suitable for the growth of many kinds of cell monolayers, and has autoclavable varieties, which is one of the most basic and widely tried mediums. However, due to its limited nutrient composition, it is not necessarily the most effective or economical medium to use for the production of specific cell culture and expression.

C. DMEM cell culture medium

DMEM is an Eagle medium modified by Dulbecco, initially designed for mouse fibroblasts. DMEM has twice the amino acid concentration and four times the vitamin concentration of MEM, with double the HCO3- and CO2 concentration for better buffering. The original formulation contained 1000 mg/L of glucose, which was later adjusted to 4500 mg/L for the growth of certain cells, which is often referred to as low and high glucose.

Low sugar is suitable for dependent adherent cell cultures, especially for tumor cell cultures with a fast growth rate and poor adhesion. High sugar is more suitable for high-density suspension cell culture, also for clonal culture with poor attachment but does not want it to be detached from its original growth point, and also for the culture of myeloma cells and transformed cells transfected with DNA in hybridoma. For example, CHO cells express the production of the hepatitis B vaccine, and CHO cells express EPO.

D. IMDM cell culture medium

Guilber and Iscove adapted Dulbecco's Medium to Iscove's Medium for the culture of erythrocytes and macrophage precursors. IMDM also promotes the growth of mouse B lymphocytes, LPS-stimulated B cells, bone marrow hematopoietic cells, T cells, and lymphoma cells. IMDM is a very nutrient-rich medium and can therefore be used for the rapid proliferation of high-density cells.

E. RPMI-1640 cell culture medium

Moore et al. developed an RPMI-1640 cell culture medium at Roswell Park Memorial Institute in 1967. It is designed for lymphocyte culture and contains BSS + 21 amino acids + 11 vitamins, etc. Now it is also used for suspension cell cultures, such as mammalian, special hematopoietic cells, normal or malignantly proliferating leukocytes, hybridoma cells, and other adult lymphocytes like K-562, HL-60, Jurkat, Daudi, IM-9, T-cell lymphoma cells, and HCT-15 epithelial cells can be used for reference.

F. HamF10 cell culture medium

Designed by Ham in 1963, it contains trace elements and can be used when serum levels are low, and is suitable for clonogenic cultures. f10 is suitable for hamster and human diploid cells and is particularly suitable for amniotic fluid cell cultures.

G. DMEM/F12 cell culture medium

Ham's F12 was designed for cloning CHO cells at low serum concentrations and is now also widely used for the analysis of clone formation rates and primary cultures. DEME F12 can also be mixed with DMEM in equal volumes to obtain a product that combines high concentration and compositional diversity, and this medium has been used for many primary cultures and is more difficult to culture cell lines. It is also often used as a base medium for serum-free cultures because of its rich nutrient content and the ability to use less serum.

H. M199 cell culture medium

A cell culture medium with defined chemical composition, M-199, designed by Morgan et al. in 1950, contains 53 components in addition to BSS and is a comprehensive medium, mainly used for chicken embryo fibroblast culture. This culture medium must be supplemented with the serum to support long-term culture. m-199 can be used to culture cells of many species of origin and can culture transfected cells. It is now widely used in virology and vaccine production.

I. McCoy5A medium

Designed by MeCoy for sarcoma cells in 1959, BSS + 40 components. It can support the growth of many kinds (such as bone marrow, skin, lung, and spleen) of primary grafts. In addition to being suitable for general primary cell culture, it is mainly used as tissue biopsy culture, some lymphocyte culture, and growth support for some difficult-to-culture cells. For example, Jensen rat sarcoma fibroblasts, human lymphocytes, HT-29, BHL-100, and other epithelial cells.

J. L15 cell culture medium

L-15 culture medium is suitable for the culture of rapidly proliferating tumor cells and is used to culture tumor cell lines in the absence of CO2. This culture medium uses a phosphate-buffered system with a further improved amino acid composition and the replacement of glucose by galactose.

How to buy cell culture media?

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