DMEM Culture Medium

Dulbecco’s Modified Eagle Medium (DME/DMEM) is an enhanced version of Basal Eagle Medium (BME). It increases amino acids and vitamins to approximately 4× the BME levels and includes additional supplements to support mammalian cells with higher metabolic demands. The original formulation, developed for embryonic mouse cells, contains 1,000 mg/L (~1 g/L) glucose; a 4,500 mg/L (~4.5 g/L) high-glucose variant was later introduced and performs better for certain cell types.

I. Key Features

• High nutrient density: Substantially elevated essential amino acids and vitamins support faster proliferation and/or higher seeding densities.
• Dual glucose options: Available in low glucose (1 g/L) and high glucose (4.5 g/L) to match cellular metabolic rates and experimental objectives.
• Flexible sub-formulas: Common variants may include or omit L-glutamine, sodium pyruvate, HEPES (buffer), and phenol red (pH indicator).
• Broad applicability: Widely used for fibroblast-like, epithelial-like, and other adherent cell lines in routine culture, transfection, and imaging.

II. Common Variants

• Low-glucose DMEM (1 g/L): Closer to lower-glucose environments; frequently used for glucose-sensitive studies or when approximating more “physiologic” glucose levels.
• High-glucose DMEM (4.5 g/L): Provides ample carbon source for rapidly proliferating or highly metabolic cells; common in high-density culture or protein expression workflows.
• ± L-glutamine / ± sodium pyruvate: Choose ready-to-use vs. add-fresh based on stability and experimental design.
• ± HEPES / varying bicarbonate: Use 5–10% CO₂ as recommended by the formulation and supplier, and set the CO₂ level based on the medium’s NaHCO₃ content; HEPES improves pH stability during open-dish work or imaging.
• ± Phenol red: Phenol-red-free versions are preferred for imaging or hormone-responsive assays to reduce background.

III. Selection & Decision Guide

• High metabolic demand / rapid growth → Prefer high glucose (4.5 g/L).
• Glucose metabolism / insulin signaling / glucotoxicity studies → Prefer low glucose (1 g/L) or a custom glucose-supplementation strategy.
• Long-term imaging / frequent dish opening → Use HEPES-buffered or otherwise enhanced buffering formulations.
• Assays sensitive to phenol red (e.g., fluorescence, hormone assays) → Choose phenol-red-free.
• Ammonia buildup or glutamine instability concerns → Use stable glutamine dipeptides (e.g., alanyl-glutamine) or add L-glutamine fresh.

IV. Typical Applications & Cell Types

• Routine adherent culture: Expansion, passaging, and banking of fibroblast-like and epithelial-like cell lines.
• Transfection/expression workflows: Enhanced nutrients and buffering help maintain cell status and improve reproducibility.
• Metabolic studies: Switching between low- and high-glucose versions or titrating carbon sources to probe metabolic adaptation and phenotypic shifts.

Note: Final selection should integrate the cell line’s datasheet, prior experience, and empirical readouts (growth curves, viability, glucose/lactate usage, pH changes, etc.).

V. DMEM-Series Media & SKU Summary (Low-Glucose / No-Glucose / High-Glucose / Hybrid)

1.DMEM – Low Glucose

D1372055

D1372060

2.DMEM/F-12 – Hybrid (Composite)

D1372074	D1372080
A1372075	D778392

3.DMEM – No Glucose

D1372065

4.DMEM – High Glucose

D1372044	D1372049
D778407	D1372045
D1372053	D1372047
D1372050	D1372046

	D1372055	D1372060	D1372074	D1372080	A1372075
Component	(mg/L)	(mg/L)	(mg/L)	(mg/L)	(mg/L)
Amino Acids
Glycine	30.0	30.0	18.75	18.75	18.75
L-Arginine hydrochloride	84.0	84.0	147.5	147.5	147.5
L-Cystine 2HCl	63.0	63.0	31.29	31.29	31.29
L-Glutamine	584.0	584.0	365.0	365.0	365.0
L-Histidine hydrochloride·H2O	42.0	42.0	31.48	31.48	31.48
L-Isoleucine	105.0	105.0	54.47	54.47	54.47
L-Leucine	105.0	105.0	59.05	59.05	59.05
L-Lysine hydrochloride	146.0	146.0	91.25	91.25	91.25
L-Methionine	30.0	30.0	17.24	17.24	17.24
L-Phenylalanine	66.0	66.0	35.48	35.48	35.48
L-Serine	42.0	42.0	26.25	26.25	26.25
L-Threonine	95.0	95.0	53.45	53.45	53.45
L-Tryptophan	16.0	16.0	9.02	9.02	9.02
L-Tyrosine disodium salt dihydrate	104.0	104.0	55.79	55.79	55.79
L-Valine	94.0	94.0	52.85	52.85	52.85
L-Alanyl-L-Glutamine	——	862.0	——	——	——
L-Alanine	——	——	4.45	4.45	4.45
L-Asparagine·H₂O	——	——	7.5	7.5	7.5
L-Aspartic acid	——	——	6.65	6.65	6.65
L-Cysteine hydrochloride·H₂O	——	——	17.56	17.56	17.56
L-Glutamic acid	——	——	7.35	7.35	7.35
L-Proline	——	——	17.25	17.25	17.25
Vitamins
Choline chloride	4.0	4.0	8.98	8.98	8.98
D-Calcium pantothenate	4.0	4.0	2.24	2.24	2.24
Folic Acid	4.0	4.0	2.65	2.65	2.65
Niacinamide	4.0	4.0	2.02	2.02	2.02
Pyridoxine hydrochloride	4.0	4.0	2.013	2.013	2.00
Riboflavin	0.4	0.4	0.219	0.219	0.219
Thiamine hydrochloride	4.0	4.0	2.17	2.17	2.17
i-Inositol	7.2	7.2	12.6	12.6	12.6
Biotin	——	——	0.0035	0.0035	0.0035
Vitamin B12	——	——	0.68	0.68	0.68
Ascorbic Acid phosphate	——	——	——	——	2.5
Inorganic Salts
Calcium Chloride (CaCl2) (anhyd.)	200.0	200.0	116.6	116.6	116.6
Ferric Nitrate (Fe(NO3)3·9H2O)	0.1	0.1	0.05	0.05	0.05
Magnesium Sulfate (MgSO4) (anhyd.)	97.67	97.67	48.84	48.84	48.84
Potassium Chloride (KCl)	400.0	400.0	311.8	311.8	311.8
Sodium Chloride (NaCl)	6400.0	6400.0	6995.5	6995.5	6995.5
Sodium Phosphate monobasic (NaH2PO4·H2O)	125.0	125.0	62.5	62.5	62.5
Sodium Bicarbonate (NaHCO3)	3700.0	3700.0	2438.0	2438.0	2438.0
Cupric sulfate (CuSO₄·5H₂O)	——	——	0.0013	0.0013	0.0013
Ferric sulfate (FeSO₄·7H₂O)	——	——	0.417	0.417	0.417
Magnesium chloride (anhydrous)	——	——	28.64	28.64	28.64
Sodium Phosphate dibasic (Na₂HPO₄, anhydrous)	——	——	71.02	71.02	71.02
Zinc sulfate (ZnSO₄·7H₂O)	——	——	0.432	0.432	0.864
Proteins
Bovine Serum Albumin	——	——	——	——	400
Human Transferrin (Holo)	——	——	——	——	7.5
Insulin Recombinant Full Chain	——	——	——	——	10
Reducing Agents
Glutathione, monosodium	——	——	——	——	1
Trace Elements
Ammonium Metavanadate	——	——	——	——	0.0003
Manganous Chloride	——	——	——	——	0.00005
Sodium Selenite	——	——	——	——	0.005
Other Components
D-Glucose (Dextrose)	1000.0	1000.0	3151.0	3151.0	3151.0
Phenol Red	15.0	15.0	——	8.1	8.1
Sodium Pyruvate	110.0	110.0	55.0	55.0	110
HEPES	5958.0	——	——	——	——
Hypoxanthine Na	——	——	2.39	2.39	2.39
Linoleic Acid	——	——	0.042	0.042	0.042
Putrescine 2HCl	——	——	0.081	0.081	0.081
Thymidine	——	——	0.365	0.365	0.365
Ethanolamine	——	——	——	——	1.9
Lipoic Acid	——	——	0.105	0.105	0.105

	D778392	D1372065	D1372044	D1372049	D778407
Component	(mg/L)	(mg/L)	(mg/L)	(mg/L)	(mg/L)
Amino Acids
Glycine	18.75	30.0	30.0	30.0	30.0
L-Arginine hydrochloride	147.5	84.0	84.0	84.0	84.0
L-Cystine 2HCl	31.29	63.0	63.0	63.0	63.0
L-Glutamine	365.0	584.0	584.0	584.0	584.0
L-Histidine hydrochloride·H2O	31.48	42.0	42.0	42.0	42.0
L-Isoleucine	54.47	105.0	105.0	105.0	105.0
L-Leucine	59.05	105.0	105.0	105.0	105.0
L-Lysine hydrochloride	91.25	146.0	146.0	146.0	146.0
L-Methionine	17.24	30.0	30.0	30.0	30.0
L-Phenylalanine	35.48	66.0	66.0	66.0	66.0
L-Serine	26.25	42.0	42.0	42.0	42.0
L-Threonine	53.45	95.0	95.0	95.0	95.0
L-Tryptophan	9.02	16.0	16.0	16.0	16.0
L-Tyrosine disodium salt dihydrate	55.79	104.0	104.0	104.0	104.0
L-Valine	52.85	94.0	94.0	94.0	94.0
L-Alanyl-L-Glutamine	——	——	——	——	——
L-Alanine	4.45	——	——	——	——
L-Asparagine·H₂O	7.5	——	——	——	——
L-Aspartic acid	6.65	——	——	——	——
L-Cysteine hydrochloride·H₂O	17.56	——	——	——	——
L-Glutamic acid	7.35	——	——	——	——
L-Proline	17.25	——	——	——	——
Vitamins
Choline chloride	8.98	4.0	4.0	4.0	4.0
D-Calcium pantothenate	2.24	4.0	4.0	4.0	4.0
Folic Acid	2.65	4.0	4.0	4.0	4.0
Niacinamide	2.02	4.0	4.0	4.0	4.0
Pyridoxine hydrochloride	2.0	4.0	4.0	4.0	4.0
Riboflavin	0.219	0.4	0.4	0.4	0.4
Thiamine hydrochloride	2.17	4.0	4.0	4.0	4.0
i-Inositol	12.6	7.2	7.2	7.2	7.2
Biotin	0.0035	——	——	——	——
Vitamin B12	0.68	——	——	——	——
Ascorbic Acid phosphate	——	——	——	——	——
Inorganic Salts
Calcium Chloride (CaCl2) (anhyd.)	116.6	200.0	200.0	200.0	200.0
Ferric Nitrate (Fe(NO3)3·9H2O)	0.05	0.1	0.1	0.1	0.1
Magnesium Sulfate (MgSO4) (anhyd.)	48.84	97.67	97.67	97.67	97.67
Potassium Chloride (KCl)	311.8	400.0	400.0	400.0	400.0
Sodium Chloride (NaCl)	6995.5	6400.0	6400.0	6400.0	6400.0
Sodium Phosphate monobasic (NaH2PO4·H2O)	62.5	125.0	125.0	125.0	125.0
Sodium Bicarbonate (NaHCO3)	1200.0	3700.0	3700.0	3700.0	3700.0
Cupric sulfate (CuSO₄·5H₂O)	0.0013	——	——	——	——
Ferric sulfate (FeSO₄·7H₂O)	0.417	——	——	——	——
Magnesium chloride (anhydrous)	28.64	——	——	——	——
Sodium Phosphate dibasic (Na₂HPO₄, anhydrous)	71.02	——	——	——	——
Zinc sulfate (ZnSO₄·7H₂O)	0.432	——	——	——	——
Proteins
Bovine Serum Albumin	——	——	——	——	——
Human Transferrin (Holo)	——	——	——	——	——
Insulin Recombinant Full Chain	——	——	——	——	——
Reducing Agents
Glutathione, monosodium	——	——	——	——	——
Trace Elements
Ammonium Metavanadate	0.0003	——	——	——	——
Manganous Chloride	0.00005	——	——	——	——
Sodium Selenite	0.005	——	——	——	——
Other Components
D-Glucose (Dextrose)	3151.0	——	4500.0	4500.0	4500.0
Phenol Red	8.1	15.0	15.0	——	15.0
Sodium Pyruvate	55.0	——	——	——	110.0
HEPES	3574.5	——	——	5958.0	——
Hypoxanthine Na	2.39	——	——	——	——
Linoleic Acid	0.042	——	——	——	——
Putrescine 2HCl	0.081	——	——	——	——
Thymidine	0.365	——	——	——	——
Ethanolamine	1.9	——	——	——	——
Lipoic Acid	0.105	——	——	——	——

	D1372045	D1372053	D1372047	D1372050	D1372046
Component	(mg/L)	(mg/L)	(mg/L)	(mg/L)	(mg/L)
Amino Acids
Glycine	30.0	30.0	30.0	30.0	30.0
L-Arginine hydrochloride	84.0	84.0	84.0	84.0	84.0
L-Cystine 2HCl	63.0	63.0	63.0	63.0	63.0
L-Glutamine	584.0	——	584.0	——	——
L-Histidine hydrochloride·H2O	42.0	42.0	42.0	42.0	42.0
L-Isoleucine	105.0	105.0	105.0	105.0	105.0
L-Leucine	105.0	105.0	105.0	105.0	105.0
L-Lysine hydrochloride	146.0	146.0	146.0	146.0	146.0
L-Methionine	30.0	30.0	30.0	30.0	30.0
L-Phenylalanine	66.0	66.0	66.0	66.0	66.0
L-Serine	42.0	42.0	42.0	42.0	42.0
L-Threonine	95.0	95.0	95.0	95.0	95.0
L-Tryptophan	16.0	16.0	16.0	16.0	16.0
L-Tyrosine disodium salt dihydrate	104.0	104.0	104.0	104.0	104.0
L-Valine	94.0	94.0	94.0	94.0	94.0
L-Alanyl-L-Glutamine	862.0	——	——	——	862.0
L-Alanine	——	——	——	——	——
L-Asparagine·H₂O	——	——	——	——	——
L-Aspartic acid	——	——	——	——	——
L-Cysteine hydrochloride·H₂O	——	——	——	——	——
L-Glutamic acid	——	——	——	——	——
L-Proline	——	——	——	——	——
Vitamins
Choline chloride	4.0	4.0	4.0	4.0	4.0
D-Calcium pantothenate	4.0	4.0	4.0	4.0	4.0
Folic Acid	4.0	4.0	4.0	4.0	4.0
Niacinamide	4.0	4.0	4.0	4.0	4.0
Pyridoxine hydrochloride	4.0	4.0	4.0	4.0	4.0
Riboflavin	0.4	0.4	0.4	0.4	0.4
Thiamine hydrochloride	4.0	4.0	4.0	4.0	4.0
i-Inositol	7.2	7.2	7.2	7.2	7.2
Biotin	——	——	——	——	——
Vitamin B12	——	——	——	——	——
Ascorbic Acid phosphate	——	——	——	——	——
Inorganic Salts
Calcium Chloride (CaCl2) (anhyd.)	200.0	——	200.0	200.0	200.0
Ferric Nitrate (Fe(NO3)3·9H2O)	0.1	0.1	0.1	0.1	0.1
Magnesium Sulfate (MgSO4) (anhyd.)	97.67	97.67	97.67	97.67	97.67
Potassium Chloride (KCl)	400.0	400.0	400.0	400.0	400.0
Sodium Chloride (NaCl)	6400.0	6400.0	6400.0	6400.0	6400.0
Sodium Phosphate monobasic (NaH2PO4·H2O)	125.0	125.0	125.0	125.0	125.0
Sodium Bicarbonate (NaHCO3)	3700.0	3700.0	3700.0	3700.0	3700.0
Cupric sulfate (CuSO₄·5H₂O)	——	——	——	——	——
Ferric sulfate (FeSO₄·7H₂O)	——	——	——	——	——
Magnesium chloride (anhydrous)	——	——	——	——	——
Sodium Phosphate dibasic (Na₂HPO₄, anhydrous)	——	——	——	——	——
Zinc sulfate (ZnSO₄·7H₂O)	——	——	——	——	——
Proteins
Bovine Serum Albumin	——	——	——	——	——
Human Transferrin (Holo)	——	——	——	——	——
Insulin Recombinant Full Chain	——	——	——	——	——
Reducing Agents
Glutathione, monosodium	——	——	——	——	——
Trace Elements
Ammonium Metavanadate	——	——	——	——	——
Manganous Chloride	——	——	——	——	——
Sodium Selenite	——	——	——	——	——
Other Components
D-Glucose (Dextrose)	4500.0	4500.0	4500.0	4500.0	4500.0
Phenol Red	15.0	15.0	15.0	15.0	15.0
Sodium Pyruvate	——	——	——	110.0	110.0
HEPES	——	——	5958.0	——	——
Hypoxanthine Na	——	——	——	——	——
Linoleic Acid	——	——	——	——	——
Putrescine 2HCl	——	——	——	——	——
Thymidine	——	——	——	——	——
Ethanolamine	——	——	——	——	——
Lipoic Acid	——	——	——	——	——

References

1. Smith, J.D., Freeman, G., Vogt, M., and Dulbecco, R., The Nucleic Acid of Polyoma. Virus, 12, 185-196 (1960).

2. Rutzky, L.P., and Pumper, R.W., Supplement to a Survey of Commercially Available Tissue Culture Media

(1970). In Vitro, 9, 468 (1974).

3. Dulbecco, R., and Freeman, G., Plaque Production by the Polyoma Virus. Virology, 8, 396-397 (1959).

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