Home Mammalian Cell CultureRPMI-1640 Media Formulation

RPMI-1640 Media Formulation

RPMI-1640 Medium Dutch Modification
RPMI-1640 Medium HEPES Modification
RPMI-1640 Medium Modified
RPMI-1640 Medium Auto-Mod™ for Autoclaving

RPMI-1640 was developed by Moore et. al. at Roswell Park Memorial Institute, hence the acronym RPMI. The formulation is based on the RPMI-1630 series of media utilizing a bicarbonate buffering system and alterations in the amounts of amino acids and vitamins. RPMI-1640 medium is commonly used for suspension cultures of mammalian and primary cells, such as HeLa cells, human peripheral blood mononuclear cells (PBMCs), human normal and neoplastic leukocytes, Jukrat cells, and more. RPMI-1640, when properly supplemented, has demonstrated wide applicability for supporting growth of many types of cultured cells, including fresh human lymphocytes in the 72-hour phytohemaglutinin (PHA) stimulation assay.

RPMI-1640 Medium Composition

Component	R0883 [1x] g/L	R1145 [10x] g/L	R1383 g/L	R1780 [1x] g/L	R2405 [1x] g/L	R6504 g/L	R8758 [1x] g/L
Inorganic Salts
Calcium Nitrate • 4H₂O	0.1	1	0.1	0.1	0.1	0.1	0.1
Magnesium Sulfate (anhydrous)	0.04884	0.4884	0.04884	0.04884	0.04884	0.04884	0.04884
Potassium Chloride	0.4	4	0.4	0.4	0.4	0.4	0.4
Sodium Bicarbonate	2	—	—	2	2	—	2
Sodium Chloride	6	60	6	6	6	6	6
Sodium Phosphate Dibasic (anhydrous)	0.8	8	0.8	0.8	0.8	0.8	0.8
Amino Acids
L-Alanyl-L-Glutamine	—	—	—	—	0.4344	—	—
L-Arginine	0.2	2	0.2	—	0.2	0.2	0.2
L-Asparagine (anhydrous)	0.05	0.5	0.05	0.05	0.05	0.05	0.05
L-Aspartic Acid	0.02	0.2	0.02	0.02	0.02	0.02	0.02
L-Cystine • 2HCl	0.0652	0.652	0.0652	0.0652	0.0652	0.0652	0.0652
L-Glutamic Acid	0.02	0.2	0.02	0.02	0.02	0.02	0.02
L-Glutamine	—	—	0.3	0.3	—	0.3	0.3
Glycine	0.01	0.1	0.01	0.01	0.01	0.01	0.01
L-Histidine	0.015	0.15	0.015	0.015	0.015	0.015	0.015
Hydroxy-L-Proline	0.02	0.2	0.02	0.02	0.02	0.02	0.02
L-Isoleucine	0.05	0.5	0.05	0.05	0.05	0.05	0.05
L-Leucine	0.05	0.5	0.05	—	0.05	0.05	0.05
L-Lysine • HCl	0.04	0.4	0.04	—	0.04	0.04	0.04
L-Methionine	0.015	0.15	0.015	0.015	0.015	0.015	0.015
L-Phenylalanine	0.015	0.15	0.015	0.015	0.015	0.015	0.015
L-Proline	0.02	0.2	0.02	0.02	0.02	0.02	0.02
L-Serine	0.03	0.3	0.03	0.03	0.03	0.03	0.03
L-Threonine	0.02	0.2	0.02	0.02	0.02	0.02	0.02
L-Tryptophan	0.005	0.05	0.005	0.005	0.005	0.005	0.005
L-Tyrosine • 2Na • 2H₂O	0.02883	0.2883	0.02883	0.02883	0.02883	0.02883	0.02883
L-Valine	0.02	0.2	0.02	0.02	0.02	0.02	0.02
Vitamins
D-Biotin	0.0002	0.002	0.0002	0.002	0.002	0.0002	0.0002
Choline Chloride	0.003	0.03	0.003	0.003	0.003	0.003	0.003
Folic Acid	0.001	—	0.001	0.001	0.001	0.001	0.001
myo-Inositol	0.035	0.35	0.035	0.035	0.035	0.035	0.035
Niacinamide	0.001	0.01	0.001	0.001	0.001	0.001	0.001
p-Aminobenzoic Acid	0.001	0.01	0.001	0.001	0.001	0.001	0.001
D-Pantothenic Acid (hemicalcium)	0.00025	0.0025	0.00025	0.00025	0.00025	0.00025	0.00025
Pyridoxine • HCl	0.001	0.01	0.001	0.001	0.001	0.001	0.001
Riboflavin	0.0002	0.002	0.0002	0.0002	0.0002	0.0002	0.0002
Thiamine • HCl	0.001	0.01	0.001	0.001	0.001	0.001	0.001
Vitamin B₁₂	0.000005	0.00005	0.000005	0.000005	0.000005	0.000005	0.000005
Other
D-Glucose	2	20	—	2	2	2	2
Glutathione (reduced)	0.001	0.01	0.001	0.001	0.001	0.001	0.001
Phenol Red • Na	0.0053	0.053	0.0053	—	0.0053	0.0053	0.0053
Add
L-Glutamine	0.3	0.3 at 1×	—	—	—	—	—
Sodium Bicarbonate	—	2.0 at 1×	2	—	—	2	—

RPMI-1640 Medium Dutch Modification

Component	R7638 [1x] g/L
Inorganic Salts
Calcium Nitrate • 4H₂O	0.1
Magnesium Sulfate (anhydrous)	0.04884
Potassium Chloride	0.4
Sodium Bicarbonate	1
Sodium Chloride	6.4
Sodium Phosphate Dibasic (anhydrous)	0.8
Amino Acids
L-Arginine	0.2
L-Asparagine (anhydrous)	0.05
L-Aspartic Acid	0.02
L-Cystine • 2HCl	0.0652
L-Glutamic Acid	0.02
Glycine	0.01
L-Histidine	0.015
Hydroxy-L-Proline	0.02
L-Isoleucine	0.05
L-Leucine	0.05
L-Lysine • HCl	0.04
L-Methionine	0.015
L-Phenylalanine	0.015
L-Proline	0.02
L-Serine	0.03
L-Threonine	0.02
L-Tryptophan	0.005
L-Tyrosine • 2Na • 2H₂O	0.02883
L-Valine	0.02
Vitamins
D-Biotin	0.0002
Choline Chloride	0.003
Folic Acid	0.001
myo-Inositol	0.035
Niacinamide	0.001
p-Aminobenzoic Acid	0.001
D-Pantothenic Acid (hemicalcium)	0.00025
Pyridoxine • HCl	0.001
Riboflavin	0.0002
Thiamine • HCl	0.001
Vitamin B₁₂	0.000005
Other
D-Glucose	2
Glutathione (reduced)	0.001
HEPES	4.77
Phenol Red • Na	0.0053
Add
L-Glutamine	0.3

RPMI-1640 Medium HEPES Modification

Component	R4130 g/L	R5886 [1x] g/L	R8005 g/L
Inorganic Salts
Calcium Nitrate • 4H₂O	0.1	0.1	0.1
Magnesium Sulfate (anhydrous)	0.04884	0.04884	0.04884
Potassium Chloride	0.4	0.4	0.4
Sodium Bicarbonate	—	2	—
Sodium Chloride	6	6	5.9
Sodium Phosphate Dibasic (anhydrous)	0.8	0.8	0.8
Amino Acids
L-Arginine	0.2	0.2	0.2
L-Asparagine (anhydrous)	0.05	0.05	0.05
L-Aspartic Acid	0.02	0.02	0.02
L-Cystine • 2HCl	0.0652	0.0652	0.0652
L-Glutamic Acid	0.02	0.02	0.02
L-Glutamine	0.3	—	0.3
Glycine	0.01	0.01	0.01
L-Histidine	0.015	0.015	0.015
Hydroxy-L-Proline	0.02	0.02	0.02
L-Isoleucine	0.05	0.05	0.05
L-Leucine	0.05	0.05	0.05
L-Lysine • HCl	0.04	0.04	0.04
L-Methionine	0.015	0.015	0.015
L-Phenylalanine	0.015	0.015	0.015
L-Proline	0.02	0.02	0.02
L-Serine	0.03	0.03	0.03
L-Threonine	0.02	0.02	0.02
L-Tryptophan	0.005	0.005	0.005
L-Tyrosine • 2Na • 2H₂O	0.02883	0.02883	0.02883
L-Valine	0.02	0.02	0.02
Vitamins
D-Biotin	0.0002	0.0002	0.0002
Choline Chloride	0.003	0.003	0.003
Folic Acid	0.001	0.001	0.001
myo-Inositol	0.035	0.035	0.035
Niacinamide	0.001	0.001	0.001
p-Aminobenzoic Acid	0.001	0.001	0.001
D-Pantothenic Acid (hemicalcium)	0.00025	0.00025	0.00025
Pyridoxine • HCl	0.001	0.001	0.001
Riboflavin	0.0002	0.0002	0.0002
Thiamine • HCl	0.001	0.001	0.001
Vitamin B₁₂	0.000005	0.000005	0.000005
Other
D-Glucose	2	2	4.5
Glutathione (reduced)	0.001	0.001	0.001
HEPES	5.96	5.96	3.5745
Phenol Red • Na	0.0053	0.0053	0.0053
Add
L-Glutamine	—	0.3	—
Sodium Bicarbonate	2	—	2

RPMI-1640 Medium Modified

Component	R7388 [1x] g/L	R7509 [1x] g/L	R7513 [1x] g/L	R8755 g/L
Inorganic Salts
Calcium Nitrate • 4H₂O	0.1	0.1	0.1	0.1
Magnesium Sulfate (anhydrous)	0.04884	0.04884	0.04884	0.04884
Potassium Chloride	0.4	0.4	0.4	0.4
Sodium Bicarbonate	—	2	2	—
Sodium Chloride	6	6	6	6
Sodium Phosphate Dibasic (anhydrous)	0.8	0.8	0.8	0.8
Amino Acids
L-Arginine	0.2	0.2	0.2	0.2
L-Asparagine (anhydrous)	0.05	0.05	0.05	0.05
L-Aspartic Acid	0.02	0.02	0.02	0.02
L-Cystine • 2HCl	0.0652	0.0652	—	0.0652
L-Glutamic Acid	0.02	0.02	0.02	0.02
L-Glutamine	0.3	—	—	0.3
Glycine	0.01	0.01	0.01	0.01
L-Histidine	0.015	0.015	0.015	0.015
Hydroxy-L-Proline	0.02	0.02	0.02	0.02
L-Isoleucine	0.05	0.05	0.05	0.05
L-Leucine	0.05	0.05	0.05	0.05
L-Lysine • HCl	0.04	0.04	0.04	0.04
L-Methionine	0.015	0.015	—	0.015
L-Phenylalanine	0.015	0.015	0.015	0.015
L-Proline	0.02	0.02	0.02	0.02
L-Serine	0.03	0.03	0.03	0.03
L-Threonine	0.02	0.02	0.02	0.02
L-Tryptophan	0.005	0.005	0.005	0.005
L-Tyrosine • 2Na • 2H₂O	0.02883	0.02883	0.02883	0.02883
L-Valine	0.02	0.02	0.02	0.02
Vitamins
D-Biotin	0.0002	0.0002	0.0002	0.0002
Choline Chloride	0.003	0.003	0.003	0.003
Folic Acid	0.001	0.001	0.001	0.001
myo-Inositol	0.035	0.035	0.035	0.035
Niacinamide	0.001	0.001	0.001	0.001
p-Aminobenzoic Acid	0.001	0.001	0.001	0.001
D-Pantothenic Acid (hemicalcium)	0.00025	0.00025	0.00025	0.00025
Pyridoxine • HCl	0.001	0.001	0.001	0.001
Riboflavin	0.0002	0.0002	0.0002	0.0002
Thiamine • HCl	0.001	0.001	0.001	0.001
Vitamin B₁₂	0.000005	0.000005	0.000005	0.000005
Other
D-Glucose	2	2	2	2
Glutathione (reduced)	0.001	0.001	0.001	0.001
HEPES	4.77	—	—	—
Phenol Red • Na	0.0053	—	0.0053	—
Add
L-Cystine • 2HCl	—	—	0.0652	—
L-Glutamine	—	0.3	0.3	—
L-Methionine	—	—	0.015	—
Sodium Bicarbonate	2	—	—	2

RPMI-1640 Medium Auto-Mod™ Modified for Autoclaving

Component	R7755 g/L
Inorganic Salts
Calcium Nitrate • 4H₂O	0.1
Magnesium Sulfate (anhydrous)	0.04884
Potassium Chloride	0.4
Sodium Chloride	6
Sodium Phosphate Dibasic (anhydrous)	0.8
Succinic Acid • 6H₂O • Na	0.1
Succinic Acid (free acid)	0.075
Amino Acids
L-Arginine	0.2
L-Asparagine (anhydrous)	0.05
L-Aspartic Acid	0.02
L-Cystine • 2HCl	0.0652
L-Glutamic Acid	0.02
Glycine	0.01
L-Histidine	0.015
Hydroxy-L-Proline	0.02
L-Isoleucine	0.05
L-Leucine	0.05
L-Lysine • HCl	0.04
L-Methionine	0.015
L-Phenylalanine	0.015
L-Proline	0.02
L-Serine	0.03
L-Threonine	0.02
L-Tryptophan	0.005
L-Tyrosine	0.02
L-Valine	0.02
Vitamins
D-Biotin	0.0002
Choline Bitartrate	0.00544
Folic Acid	0.001
myo-Inositol	0.035
Niacinamide	0.001
p-Aminobenzoic Acid	0.001
D-Pantothenic Acid (hemicalcium)	0.00025
Pyridoxine • HCl	0.001
Riboflavin	0.0002
Thiamine • HCl	0.001
Vitamin B₁₂	0.000005
Other
D-Glucose	2
Glutathione (reduced)	0.001
Phenol Red • Na	0.00318
Add
L-Glutamine	0.3
Sodium Bicarbonate	2

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