This article is missing information about different salts (Hanks, Earle), actual composition.(November 2018) |
Minimal essential medium (MEM) is a synthetic cell culture medium developed by Harry Eagle first published in 1959 [1] in Science that can be used to maintain cells in tissue culture. It is based on 6 salts and glucose described in Earle's salts in 1934: (calcium chloride, potassium chloride, magnesium sulfate, sodium chloride, sodium phosphate and sodium bicarbonate), supplemented with 13 essential amino acids, and 8 vitamins: thiamine (vitamin B1), riboflavin (vitamin B2), nicotinamide (vitamin B3), pantothenic acid (vitamin B5), pyrodoxine (vitamin B6), folic acid (vitamin B9), choline, and myo-inositol (originally known as vitamin B8).
Many variations of this medium have been developed, mostly adding additional vitamins, amino acids, and/or other nutrients. [2]
Eagle developed his earlier "Basal Medium Eagle" (BME) in 1955–1957 on mouse L cells [3] and human HeLa cells, [4] with 13 essential amino acids and 9 vitamins added. BME contains biotin (vitamin B7), which Eagle later found to be superfluous. His 1959 "minimal essential medium" doubles the amount of many amino acids to "conform more closely to the protein composition of cultured human cells. This permits the cultures to be kept for somewhat longer periods without refeeding". [1]
DMEM (Dulbecco's modified Eagle's medium) was originally suggested as Eagle's medium with a 'fourfold concentration of amino acids and vitamins' by Renato Dulbecco and G. Freeman published in 1959. [5] The commercial versions of this medium have additional modifications, see an example in the table below. [6]
α-MEM (minimal essential medium Eagle – alpha modification) is a medium based on MEM published in 1971 by Clifford P. Stanners and colleagues. [7] It contains more non-essential amino acids, sodium pyruvate, and vitamins (ascorbic acid (vitamin C), biotin, and cyanocobalamin) compared with MEM. It can also come with lipoic acid and nucleosides. [8] [9]
Glasgow's MEM (Glasgow minimal essential medium) is yet another modification, prepared by Ian MacPherson and Michael Stoker. [10]
One liter of each medium contains (in milligrams):
Medium | BME [11] | MEM [12] | α-MEMa [13] | DMEM [6] |
Glycine | 50 | 30 | ||
L-alanine | 25 | |||
L-arginine hydrochloride | 21 | 126 | 126 | 84 |
L-asparagine-H2O | 50 | |||
L-aspartic acid | 30 | |||
L-cysteine hydrochloride-H2O | 100 | |||
L-cystine 2HCl | 16 | 31 | 31 | 63 |
L-glutamic acid | 75 | |||
L-glutamine | 292 | 292 | 292 | 584 |
L-histidine | 8 | 31 | 42 | |
L-histidine hydrochloride-H2O | 42 | 42 | ||
L-isoleucine | 26 | 52 | 52 | 105 |
L-leucine | 26 | 52 | 52 | 105 |
L-lysine hydrochloride | 36.47 | 73 | 73 | 146 |
L-methionine | 7.5 | 15 | 15 | 30 |
L-phenylalanine | 16.5 | 32 | 32 | 66 |
L-proline | 40 | |||
L-serine | 25 | 42 | ||
L-threonine | 24 | 48 | 48 | 95 |
L-tryptophan | 4 | 10 | 10 | 16 |
L-tyrosine disodium salt dihydrate | 26 | 52 | 52 | 104 |
L-valine | 23.5 | 46 | 46 | 94 |
Ascorbic acid | 50 | |||
Biotin | 1 | 0.1 | ||
Choline chloride | 1 | 1 | 1 | 4 |
D-calcium pantothenate | 1 | 1 | 1 | 4 |
Folic acid | 1 | 1 | 1 | 4 |
Niacinamide | 1 | 1 | 1 | 4 |
Pyridoxal hydrochloride | 1 | 1 | 1 | 4 |
Riboflavin | 0.1 | 0.1 | 0.1 | 0.4 |
Thiamine hydrochloride | 1 | 1 | 1 | 4 |
Vitamin B12 | 1.36 | |||
i-Inositol | 2 | 2 | 2 | 7.2 |
Calcium chloride (CaCl2, anhyd.) | 200 | 200 | 200 | 200 |
Ferric nitrate (Fe(NO3)3·9H2O) | 0.1 | |||
Magnesium sulfate (MgSO4, anhyd.) | 97.67 | 97.67 | 97.67 | 97.67 |
Potassium chloride (KCl) | 400 | 400 | 400 | 400 |
Sodium bicarbonate (NaHCO3) | 2200 | 2200 | 2200 | 3700 |
Sodium chloride (NaCl) | 6800 | 6800 | 6800 | 6400 |
Sodium phosphate monobasic (NaH2PO4-H2O) | 140 | 140 | 140 | 125 |
D-Glucose (dextrose) | 1000 | 1000 | 1000 | 1000 |
Lipoic acid | 0.2 | |||
Phenol red | 10 | 10 | 10 | 15 |
Sodium pyruvate | 110 | 110 |
The citric acid cycle—also known as the Krebs cycle, Szent–Györgyi–Krebs cycle or the TCA cycle (tricarboxylic acid cycle)—is a series of biochemical reactions to release the energy stored in nutrients through the oxidation of acetyl-CoA derived from carbohydrates, fats, and proteins. The chemical energy released is available under the form of ATP. The Krebs cycle is used by organisms that respire (as opposed to organisms that ferment) to generate energy, either by anaerobic respiration or aerobic respiration. In addition, the cycle provides precursors of certain amino acids, as well as the reducing agent NADH, that are used in numerous other reactions. Its central importance to many biochemical pathways suggests that it was one of the earliest components of metabolism. Even though it is branded as a "cycle", it is not necessary for metabolites to follow only one specific route; at least three alternative segments of the citric acid cycle have been recognized.
α-Ketoglutaric acid is a dicarboxylic acid, i.e., a short-chain fatty acid containing two carboxyl groups with C, O, and H standing for carbon, oxygen, and hydrogen, respectively. However, almost all animal tissues and extracellular fluids have a pH above 7. At these basic pH levels α-ketoglutaric acid exists almost exclusively as its conjugate base. That is, it has two negative electric charges due to its release of positively charged hydrogen from both of its now negatively charged carboxy groups, CO−2. This double negatively charge molecule is referred to as α-ketoglutarate or 2-oxoglutarate.
Thiamine, also known as thiamin and vitamin B1, is a vitamin, an essential micronutrient for humans and animals. It is found in food and commercially synthesized to be a dietary supplement or medication. Phosphorylated forms of thiamine are required for some metabolic reactions, including the breakdown of glucose and amino acids.
Pantothenic acid (vitamin B5) is a B vitamin and an essential nutrient. All animals need pantothenic acid in order to synthesize coenzyme A (CoA), which is essential for cellular energy production and for the synthesis and degradation of proteins, carbohydrates, and fats.
Biotin (also known as vitamin B7 or vitamin H) is one of the B vitamins. It is involved in a wide range of metabolic processes, both in humans and in other organisms, primarily related to the utilization of fats, carbohydrates, and amino acids. The name biotin, borrowed from the German Biotin, derives from the Ancient Greek word βίοτος (bíotos; 'life') and the suffix "-in" (a suffix used in chemistry usually to indicate 'forming'). Biotin appears as a white, needle-like crystalline solid.
Alanine, or α-alanine, is an α-amino acid that is used in the biosynthesis of proteins. It contains an amine group and a carboxylic acid group, both attached to the central carbon atom which also carries a methyl group side chain. Consequently it is classified as a nonpolar, aliphatic α-amino acid. Under biological conditions, it exists in its zwitterionic form with its amine group protonated and its carboxyl group deprotonated. It is non-essential to humans as it can be synthesized metabolically and does not need to be present in the diet. It is encoded by all codons starting with GC.
Threonine is an amino acid that is used in the biosynthesis of proteins. It contains an α-amino group, a carboxyl group, and a side chain containing a hydroxyl group, making it a polar, uncharged amino acid. It is essential in humans, meaning the body cannot synthesize it: it must be obtained from the diet. Threonine is synthesized from aspartate in bacteria such as E. coli. It is encoded by all the codons starting AC.
A nutrient is a substance used by an organism to survive, grow, and reproduce. The requirement for dietary nutrient intake applies to animals, plants, fungi, and protists. Nutrients can be incorporated into cells for metabolic purposes or excreted by cells to create non-cellular structures, such as hair, scales, feathers, or exoskeletons. Some nutrients can be metabolically converted to smaller molecules in the process of releasing energy, such as for carbohydrates, lipids, proteins, and fermentation products, leading to end-products of water and carbon dioxide. All organisms require water. Essential nutrients for animals are the energy sources, some of the amino acids that are combined to create proteins, a subset of fatty acids, vitamins and certain minerals. Plants require more diverse minerals absorbed through roots, plus carbon dioxide and oxygen absorbed through leaves. Fungi live on dead or living organic matter and meet nutrient needs from their host.
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RPMI 1640, simply known as RPMI medium, is a cell culture medium commonly used to culture mammalian cells. RPMI 1640 was developed by George E. Moore, Robert E. Gerner, and H. Addison Franklin in 1966 at Roswell Park Comprehensive Cancer Center, from where it derives its name. A modification of McCoy′s 5A medium, it was originally formulated to support lymphoblastoid cells in suspension cultures, but can also support a wide variety of adherent cells.
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