Medium 199 Formulation

In 1950, J.F. Morgan conducted experiments with various combinations of vitamins, amino acids, and other factors to develop the novel culture media Medium 199. When properly supplemented, Medium 199 (M199) has broad species applicability, particularly for cultivation of non-transformed cells. It is widely used in virology, vaccine production, and in vitro cultivation of primary explants of mouse pancreatic epithelial and rat lens tissues.

Component	M4530 [1x] g/L	M5017 g/L	M7528 [1x] g/L	M7653 [1x] g/L	M9163 [10x] g/L
Inorganic Salts
Calcium Chloride	0.2	0.2	0.2	0.1396	1.396
Ferric Nitrate • 9H2O	0.00072	0.00072	0.00072	0.00072	0.0072
Magnesium Sulfate (anhydrous)	0.09767	0.09767	0.09767	0.09767	0.9767
Potassium Chloride	0.4	0.4	0.4	0.4	4
Potassium Phosphate Monobasic	—	—	—	0.06	0.6
Sodium Acetate (anhydrous)	0.05	0.05	0.05	0.05	0.5
Sodium Bicarbonate	2.2	—	2.2	0.35	—
Sodium Chloride	6.8	6.8	6	8	80
Sodium Phosphate Dibasic (anhydrous)	—	—	—	0.04788	0.4788
Sodium Phosphate Monobasic (anhydrous)	0.122	0.122	0.122	—	—
Amino Acids
L-Alanine	0.025	0.025	0.025	0.025	0.25
L-Arginine • HCl	0.07	0.07	0.07	0.07	0.7
L-Aspartic Acid	0.03	0.03	0.03	0.03	0.3
L-Cysteine • HCl • H2O	0.00011	0.00011	0.00011	0.00011	0.0011
L-Cystine • 2HCl	0.026	0.026	0.026	0.026	0.26
L-Glutamic Acid	0.0668	0.0668	0.0668	0.0668	0.668
L-Glutamine	0.1	0.1	—	—	—
Glycine	0.05	0.05	0.05	0.05	0.5
L-Histidine • HCl • H2O	0.02188	0.02188	0.02188	0.02188	0.2188
Hydroxy-L-Proline	0.01	0.01	0.01	0.01	0.1
L-Isoleucine	0.02	0.02	0.02	0.02	0.2
L-Leucine	0.06	0.06	0.06	0.06	0.6
L-Lysine • HCl	0.07	0.07	0.07	0.07	0.7
L-Methionine	0.015	0.015	0.015	0.015	0.15
L-Phenylalanine	0.025	0.025	0.025	0.025	0.25
L-Proline	0.04	0.04	0.04	0.04	0.4
L-Serine	0.025	0.025	0.025	0.025	0.25
L-Threonine	0.03	0.03	0.03	0.03	0.3
L-Tryptophan	0.01	0.01	0.01	0.01	0.1
L-Tyrosine • 2Na • 2H2O	0.05766	0.05766	0.05766	0.05766	0.5766
L-Valine	0.025	0.025	0.025	0.025	0.25
Vitamins
Ascorbic Acid • Na	0.0000566	0.0000566	0.0000566	0.0000566	0.000566
D-Biotin	0.00001	0.00001	0.00001	0.00001	0.0001
Calciferol	0.0001	0.0001	0.0001	0.0001	0.001
Choline Chloride	0.0005	0.0005	0.0005	0.0005	0.005
Folic Acid	0.00001	0.00001	0.00001	0.00001	0.0001
Menadione (sodium bisulfite)	0.000016	0.000016	0.000016	0.000016	0.00016
myo-Inositol	0.00005	0.00005	0.00005	0.00005	0.0005
Niacinamide	0.000025	0.000025	0.000025	0.000025	0.00025
Nicotinic Acid	0.000025	0.000025	0.000025	0.000025	0.00025
p-Aminobenzoic Acid	0.00005	0.00005	0.00005	0.00005	0.0005
D-Pantothenic Acid (hemicalcium)	0.00001	0.00001	0.00001	0.00001	0.0001
Pyridoxal • HCl	0.000025	0.000025	0.000025	0.000025	0.00025
Pyridoxine • HCl	0.000025	0.000025	0.000025	0.000025	0.00025
Retinol Acetate	0.00014	0.00014	0.00014	0.00014	0.0014
Riboflavin	0.00001	0.00001	0.00001	0.00001	0.0001
DL-α-Tocopherol Phosphate • Na	0.00001	0.00001	0.00001	0.00001	0.0001
Thiamine • HCl	0.00001	0.00001	0.00001	0.00001	0.0001
Other
Adenine Sulfate	0.01	0.01	0.01	0.01	0.1
Adenosine Triphosphate • 2Na	0.001	0.001	0.001	0.001	0.01
Adenosine Monophosphate • Na	0.0002385	0.0002385	0.0002385	0.0002385	0.002385
Cholesterol	0.0002	0.0002	0.0002	0.0002	0.002
Deoxyribose	0.0005	0.0005	0.0005	0.0005	0.005
Glucose	1	1	1	1	10
Glutathione (reduced)	0.00005	0.00005	0.00005	0.00005	0.0005
Guanine • HCl	0.0003	0.0003	0.0003	0.0003	0.003
HEPES	—	—	5.958	—	—
Hypoxanthine	0.0003	0.0003	0.0003	0.0003	0.003
Phenol Red • Na	0.0213	0.0213	0.0213	0.0213	0.0213
Polyoxyethylenesorbitan Monooleate (TWEEN 80)	0.02	0.02	0.02	0.02	0.2
Ribose	0.0005	0.0005	0.0005	0.0005	0.005
Thymine	0.0003	0.0003	0.0003	0.0003	0.003
Uracil	0.0003	0.0003	0.0003	0.0003	—
Xanthine • Na	0.000344	0.000344	0.000344	0.000344	—
Add
L-Glutamine	—	—	0.1	1	0.1 at 1×
Sodium Bicarbonate	—	2.2	—	—	0.35 at 1×

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