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M2851-01 Medium 199 w/ Earle’s, L-Glutamine, w/o Glucose (Powder, TC 199)

Specifications
References
Grade
Cell Culture Grade
EU Commodity Code
38210000
Shipping Temp
RT
Storage Temp
RT/4°C

In 1950, Morgan, Morton and Parker described one of the first totally defined media that did not depend largely on animal products or extracts as nutritive sources. This medium supported the growth of primary chick embryo heart cells. It has since become known as Medium 199. Its current use is with the addition of serum in virus and vaccine production, and in the culture of many non-transformed cells. It is also used in combination with less complex media. Medium 199 modified with Earle's Balanced Salts is designed for use with cells maintained in ambient (non-CO2) atmospheric conditions. M2852 differs from the original formulation by containing all L-amino acids. The original formulation contained 10 DL-amino acids, which are less biologically active on a per gram basis than the corresponding L-amino acids.

The search for a synthetic medium to replace serum for maintaining cells in vitro began in the late nineteenth century and continues to this day. Ringer, Locks and Tyrode substituted physiological salt solutions augmented with glucose for serum and thereby laid the foundation for the development of defined media. As biochemical and analytical techniques have improved, more of the components in serum such as vitamins, hormones, and amino acids have been identified and incorporated into physiological salt solutions, reducing, and in some cases eliminating the concentrations of animal sera required as a medium supplement. Although there have been many modifications to the original formulas in efforts to produce fully defined media, salt solutions still play an important role in tissue culture. A salt solution's basic function, to maintain the pH and osmotic balance in the medium and to provide the cells with water and essential inorganic ions, is as valuable today as when it was first developed a century ago. Earle’s Balanced Salts selection is strongly influenced by 1. type of cell, 2. type of culture monolayer, suspension, clonal, and 3. degree of chemical definition necessary. It is important to review the literature for recommendations concerning medium, supplementation and physiological parameters required for a specific cell line.
Appearance
Peach, homogeneous, free flowing powder
Solubility
Yellow, clear, complete
pH
As Reported
Endotoxin
≤1EU/ml
Directions per Liter
Dissolve 8.9g in 800-900ml of ddH2O stirring gently until completely solubilized. If required add sodium bicarbonate. Adjust pH of the medium to the desired level. Add additional water to bring the solution to1L. Add carbohydrate as required. Filter-sterilize using a 0.22 micron membrane filter. Aliquot into sterile containers. Do not autoclave. Contains heat-labile compounds that can be damaged with autoclaving.
Storage and Stability
Store powdered media at RT. Stable for 12 months after receipt. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept at 4°C and used within a short period of time.
Media Formulation
Components shown as g/liter
Inorganic Salts
Calcium Chloride•2H2O0.265
Ferric Nitrate •9H2O0.00072
MagnesiumSulfate0.09767
Potassium Chloride0.4
Sodium Acetate0.05
Sodium Chloride6.8
Sodium Phosphate Monobasic0.122
Amino Acids
L-Alanine0.05
L-Arginine•HCl0.07
L-Aspartic Acid0.06
L-Cysteine•HCL•H2O0.00011
L-Cystine•2HCl0.026
L-Glutamic Acid0.1336
L-Glutamine0.1
Glycine0.05
L-Histidine•HCl•H2O0.02188
Hydroxy-L-proline0.01
L-Isoleucine0.04
L-Leucine0.12
L-Lysine•HCl0.07
L-Methionine0.03
L-Phenylalanine0.05
L-Proline0.04
L-Serine0.05
L-Threonine0.06
L-Tryptophan0.02
L-Tyrosine•2Na•2H2O0.05766
L-Valine0.05
Vitamins
P-Aminobenzoic Acid0.00005
Ascorbic Acid•Na0.0000566
D-Biotin0.00001
Choline Chloride0.0005
Cocarboxylase0.00001
Calciferol0.0001
myo-Inositol0.00005
Menadione, Sodium Bisulfite0.000016
Nicotinic Acid0.000025
Niacinamide0.000025
D-Pantothenic Acid•Ca0.00001
Pyridoxal•HCl0.000025
Pyridoxine•HCl0.000025
Riboflavin0.00001
Thiamine•HCl0.00001
DL-a-Tocopherol Phosphate•2Na0.00001
Retinol Acetate0.00014
Other
Adenine Sulfate0.01
Adenosine Monophosphate0.0002385
Adenosine Triphosphate•2Na0.001
Cholesterol0.0002
2-Deoxy-D-Ribose0.0005
D-GlucoseAbsent
Glutathione0.00005
Guanine•HCl0.0003
Hypoxanthine0.0003
Phenol Red ,Sodium0.0213
Tween®800.02
D-Ribose0.0005
Thymine0.0003
Uracil0.0003
Xanthine•Na0.000344
Total:8.9g/liter
References
Langston, W. et al., (2011) Free Radical Biology and Medicine doi:10.1016/j.freeradbiomed.2011.08.004 1. Morgan, J.F., Morton, H.J. and Parker, R.C. Proc. Soc. Exp. Biol. Med. 73:1-8 (1950) 2. Morgan, J.F., Campbell, E. and Morton, H.J. J.N.C.I. 16:2, 557-567 (1955)
USBio References
US Biological application reference: Langston, W. et al., (2011) Free Radical Biology and Medicine doi:10.1016/j.freeradbiomed.2011.08.004
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