FluxMPS™ Glasgow's Minimum Essential Medium (GMEM), High Glucose with 50mM HEPES: 2X Liquid
FluxMPS™ Glasgow's Minimum Essential Medium (GMEM), High Glucose with 50mM HEPES: 2X Liquid
Double-strength (2X) GMEM High Glucose with 50 mM HEPES — 9.0 g/L glucose in the concentrate, 4.5 g/L at 1X, with no tryptose phosphate broth or NEAA — for soft-agar, plaque-overlay, amino-acid dropout and other defined or semisolid cultures, ultra-filtered through a quadruple-stage 0.1 µm / 0.04 µm filtration system. With a sodium bicarbonate + 50 mM HEPES dual buffer, it is approximately 5× cleaner than conventional 0.22 µm-filtered media by particulate count.
- 2X concentrate: mix with an equal volume of melted agar or agarose, or another compatible solution, for a final 1X medium
- No tryptose phosphate broth or NEAA, for a more defined basal medium you supplement yourself
- 50 mM HEPES (25 mM at 1X) for CO₂-independent pH stability during handling
- Quadruple-stage filtration: 0.1 µm membrane filtered twice, 0.04 µm membrane filtered twice
- 0.04 µm nano-filtration final polish acts as a mycoplasma barrier
- Endotoxin < 0.05 EU/mL (USP <85> BET)
- Formulation: [+] 9.0 g/L Glucose, [+] L-Glutamine, [+] Sodium Bicarbonate, [+] Phenol Red, [+] 50mM HEPES; [−] NEAA, [−] Tryptose Phosphate Broth
- Ultrapure Type 1 water (18.2 MΩ·cm); ISO Class 5 (Class 100) aseptic fill in Totowa, NJ
- Custom formulations available — pH, glucose, HEPES, salts, and nutrient composition on request
- D-Glucose9,000 mg/L
- L-Glutamine584 mg/L
- Sodium PyruvateNot included
- BufferNaHCO₃ 5,500 mg/L + 50 mM HEPES
- Phenol Red30 mg/L
- AppearanceRed
- pH7.4 (± 0.04)
- Endotoxin< 0.05 EU/mL
- Filtration0.1 µm ×2 + 0.04 µm ×2
- Format2X liquid, 500 mL
Engineered where standard media fails
Conventional GMEM is sterile-filtered once at 0.22 µm. That pore size can pass mycoplasma and leaves subvisible particulates that settle in microchannels, foul membranes and hydrogels, and add noise to optical readouts.[11,12] FluxMPS™ GMEM keeps the Glasgow formulation and removes that failure mode.
Microchannel-safe purity
A 0.04 µm final filter and USP <788> particulate control keep subvisible debris out of chips, tubing, membranes and matrices.
Total metabolic control
Defined 9,000 mg/L D-glucose, 584 mg/L L-glutamine and no sodium pyruvate in the 2X (half that at 1X).
Ultrapure-grade water
Made with Ultrapure Type 1 water (18.2 MΩ·cm) and released at < 0.05 EU/mL endotoxin by USP <85>.
Low background for imaging
Particle-depleted medium reduces scatter and debris artifacts in colony counting, confocal imaging and on-chip sensing.
Matrix-ready 2X strength
31 components at double strength, so melted agar, agarose, supplements or cells can be added 1:1 without diluting the medium below 1X.
Customization on demand
pH, glucose, HEPES, salts, and nutrient composition on request — contact support@diagnocine.com.
Quadruple-stage filtration system
Every lot of DCP-GMEMH-JN2X is membrane filtered at 0.1 µm twice and 0.04 µm twice — a ready-to-dilute 2X GMEM polished to a 0.04 µm final pore size, a purity level single-pass 0.22 µm media cannot reach.
-
01
0.1 µmPre-filtration I
Removes large particulates and aggregates and extends the life of the downstream filters.
-
02
0.04 µmPre-filtration II
Retains fine particulates, bacteria and mycoplasma.
-
03
0.1 µmSterile-filtration I
Second-pass redundancy that catches anything shed or bypassed upstream.
-
04
0.04 µmSterile-filtration II — Final Polish
Final nano-polish immediately before ISO Class 5 aseptic fill.
Performance vs. conventional media
By particulate count, FluxMPS™ GMEM is approximately 5× cleaner than conventional 0.22 µm-filtered media, lowering clog risk in narrow channels and long perfusion runs.
© Diagnocine® — DCP-GMEMH-JN2X
Where FluxMPS™ GMEM fits
Glasgow's Minimum Essential Medium (GMEM) is a modified Basal Medium Eagle (BME) formulation.[2] Macpherson and Stoker originally added tryptose phosphate broth and doubled BME's amino acids and vitamins to grow BHK-21 clone 13 cells.[1,3] This version contains no tryptose phosphate broth and no added non-essential amino acids. That makes it a more defined basal medium that researchers can supplement to suit their own cell systems.
GMEM has no proteins, lipids, or growth factors, so it must be supplemented for cell growth. It uses a sodium bicarbonate buffer and needs a 5–10% CO₂ atmosphere to keep physiological pH. Users should check the literature for supplements and growth requirements specific to their cell lines.
GMEM 2X, Without Tryptose Phosphate Broth and NEAA, is a double-strength version of the 1X basal formulation. It's meant to be combined with an equal volume of a compatible solution so the basal ingredients end up at 1X. This is the standard approach for agar and agarose cultures, where 2X medium is mixed 1:1 with melted agar or agarose.[4] Because the formulation contains no TPB or NEAA, researchers can add their own supplements, test compounds, cells, or gelling agents to the 2X medium while keeping the final basal concentration under control. Final pH, osmolality, buffering, supplementation, and gel compatibility have to be confirmed for each research system. The 50 mM HEPES (25 mM at 1X) adds CO₂-independent buffering for handling outside the incubator.[6]
Why omit TPB and NEAA?
- Without TPB: TPB contains tryptose, a peptone made by enzymatically digesting the milk protein casein. Leaving it out removes an undefined, hydrolysate-derived source of nutrients and survival factors.
- Without NEAA: Standard MEM NEAA supplement supplies seven amino acids: glycine, L-alanine, L-asparagine, L-aspartic acid, L-glutamic acid, L-proline, and L-serine. Researchers who omit them can add back selected amino acids at controlled levels.
It is useful for:
- Defined or serum-free medium development
- Amino-acid dropout and add-back studies
- Serine/glycine and nutrient-metabolism research
- BHK-21 and kidney-cell research
- Stem-cell differentiation research
- Soft agar and anchorage-independent growth assays
- Virology plaque or overlay assays
- Customized research media
Automated Bioreactors & Robotics
For automated bioreactors, liquid-handling robots and closed perfusion loops with fine valves and in-line sensors, an optional 10 nm (0.01 µm) ultra-filtered grade of this medium is available.
- Total Particulate Exclusion: 0.01 µm filtration removes virtually all particulates down to the nanometer scale.
- Valve & Sensor Protection: keeps micro-valves, pumps and optical or electrochemical sensors free of deposits.
- Extended Perfusion Stability: supports long, uninterrupted perfusion runs with fewer line changes.
Inquiry Required: the 0.01 µm grade is made on request — email support@diagnocine.com with your catalog number and volume.
Soft-Agar Colony Assays
Mix 1:1 with molten agar or agarose for anchorage-independent growth assays at 1X medium strength.[4]
Plaque & Overlay Assays
Mix 1:1 with agarose or methylcellulose for plaque and overlay assays at 1X medium strength; BHK-21 cells are a classic virology host.[3]
Stem-Cell Differentiation
GMEM is a common basal medium in embryonic stem-cell self-renewal and differentiation research when supplemented appropriately.[5]
Amino-Acid Dropout & Add-Back
No NEAA, so glycine, serine, proline and other non-essential amino acids can be added back at controlled levels for serine/glycine and nutrient-metabolism studies.
Organ-on-a-Chip & MPS
Microchannel-safe medium for perfused organ-, tissue- and body-on-a-chip platforms.[8,9,10]
Microscopy & Optical Sensing
Fewer particles means cleaner fields for colony counting, confocal imaging and on-chip biosensors.
Specifications & quality control
Release specifications for DCP-GMEMH-JN2X. Lot-specific values are reported on the Certificate of Analysis.
| Parameter | Specification |
|---|---|
| Formulation | [+] 9.0 g/L Glucose, [+] L-Glutamine, [+] Sodium Bicarbonate, [+] Phenol Red, [+] 50mM HEPES; [−] NEAA, [−] Tryptose Phosphate Broth |
| Concentration | 2X (dilute 1:1 to 1X) |
| Appearance | Red |
| pH | 7.4 (± 0.04) USP <791> |
| Osmolality | Measured per lot; reported on CoA USP <785> |
| D-Glucose | 9,000 mg/L (9.0 g/L; 4.5 g/L at 1X) |
| L-Glutamine | 584 mg/L |
| Sodium Pyruvate | Not included |
| Sodium Bicarbonate | 5,500 mg/L |
| Phenol Red | 30 mg/L |
| HEPES | 11,916 mg/L (50 mM; 25 mM at 1X) |
| NEAA | Not included |
| Tryptose phosphate broth | Not included |
| Parameter | Specification |
|---|---|
| Endotoxin | < 0.05 EU/mL USP <85> BET |
| Sterility | No bacterial or fungal growth after 14 days of incubation USP <71> |
| Mycoplasma | 0.04 µm barrier filtration USP <63> equiv. |
| Particulate ≥10 µm | Meets limits USP <788> M2 |
| Particulate ≥25 µm | Meets limits USP <788> M2 |
| Water purity | Ultrapure Type 1, 18.2 MΩ·cm |
| Manufacturing std. | ISO 13485:2016 ISO |
| Fill environment | ISO Class 5 (Class 100) ISO |
| Parameter | Specification |
|---|---|
| Storage temperature | 2–8 °C, protected from light |
| Freeze-thaw | Do not freeze |
| Shelf life | 12 months; use before the expiry date on the product label |
| Shipping condition | Ice Pack |
| CO₂ requirement | CO₂-controlled incubator (5–10% CO₂) after dilution to 1X; sodium bicarbonate 5,500 mg/L in the 2X, 2,750 mg/L at 1X; 50 mM HEPES (25 mM at 1X) steadies pH during handling |
| Parameter | Specification |
|---|---|
| Raw material grade | High-purity, cell-culture-tested components |
| Traceability | Lot-traceable; CoA per lot |
| Manufacturing QMS | ISO 13485:2016 ISO |
| Regulatory alignment | 21 CFR Part 820 (cGMP) aligned |
| Production method | Micro-batch 2X liquid, quadruple-stage filtered; aseptic fill and finish in Totowa, NJ |
| Intended use | For Research Use Only (RUO) |
Full composition (mg/L)
DCP-GMEMH-JN2X is 2X GMEM High Glucose with 9.0 g/L glucose, L-glutamine, sodium bicarbonate, Phenol Red and 50 mM HEPES. It does not contain non-essential amino acids (NEAA) nor tryptose phosphate broth. Users are advised to review the literature for recommendations regarding medium supplementation and physiological growth requirements specific for different cell lines. All 31 components are listed below at 2X strength; values are per-lot release basis and are halved after 1:1 dilution.
| Component | CAS Number | mg/L |
|---|---|---|
| INORGANIC SALTS | ||
| Calcium chloride dihydrate | 10035-04-8 | 530.000 |
| Ferric nitrate nonahydrate | 7782-61-8 | 0.200 |
| Magnesium sulphate anhydrous | 7487-88-9 | 195.440 |
| Potassium chloride | 7447-40-7 | 800.000 |
| Sodium bicarbonate | 144-55-8 | 5500.000 |
| Sodium chloride | 7647-14-5 | 12800.000 |
| Sodium dihydrogen phosphate anhydrous | 7558-80-7 | 218.000 |
| Component | CAS Number | mg/L |
|---|---|---|
| AMINO ACIDS | ||
| L-Arginine hydrochloride | 1119-34-2 | 84.000 |
| L-Cystine | 56-89-3 | 48.000 |
| L-Glutamine | 56-85-9 | 584.000 |
| L-Histidine hydrochloride | 5934-29-2 | 42.000 |
| L-Isoleucine | 73-32-5 | 104.800 |
| L-Leucine | 61-90-5 | 104.800 |
| L-Lysine hydrochloride | 657-27-2 | 146.200 |
| L-Methionine | 63-68-3 | 30.000 |
| L-Phenylalanine | 63-91-2 | 66.000 |
| L-Threonine | 72-19-5 | 95.200 |
| L-Tryptophan | 73-22-3 | 16.000 |
| L-Tyrosine disodium salt | 69847-45-6 | 104.000 |
| L-Valine | 72-18-4 | 93.600 |
| Component | CAS Number | mg/L |
|---|---|---|
| VITAMINS | ||
| Choline chloride | 67-48-1 | 4.000 |
| D-Ca-Pantothenate | 137-08-6 | 4.000 |
| Folic acid | 59-30-3 | 4.000 |
| Nicotinamide | 98-92-0 | 4.000 |
| Pyridoxal hydrochloride | 65-22-5 | 4.000 |
| Riboflavin | 83-88-5 | 0.400 |
| Thiamine hydrochloride | 67-03-8 | 4.000 |
| i-Inositol | 87-89-8 | 7.200 |
| OTHERS | ||
| D-Glucose | 50-99-7 | 9000.000 |
| Phenol Red | 34487-61-1 | 30.000 |
| HEPES | 7365-45-9 | 11916.000 |
Manufacturing & compliance
Manufactured under an ISO 13485:2016 quality management system; aseptic fill and finish in Totowa, NJ, with every lot released against compendial test methods.
ISO 13485:2016 QMS
Documented, audited quality management system aligned with 21 CFR Part 820 (cGMP).
Ultrapure Type 1 Water
18.2 MΩ·cm water as the formulation base for low ionic and endotoxin background.
ISO Class 5 Fill & Finish
Aseptic filling in an ISO Class 5 (Class 100) environment in Totowa, NJ, right after the final 0.04 µm polish.
Micro-Batch Precision
Small, tightly controlled batches for consistent composition and pH from lot to lot.
Endotoxin — USP <85> BET
Released at < 0.05 EU/mL.
Particulate — USP <788> Method 2
Subvisible particles ≥10 µm and ≥25 µm within compendial limits.
Osmolality — USP <785>
Measured per lot and reported on the CoA.
Documentation / CoA
Appearance, pH 7.4 (± 0.04), 14-day sterility, endotoxin and osmolality per lot.
How DCP-GMEMH-JN2X compares
Same GMEM chemistry, different purity class. Conventional columns describe typical single-pass 0.22 µm-filtered catalog media.
| Parameter | DCP-GMEMH-JN2X (FluxMPS™) | Conventional GMEM High Glucose, 1X (0.22 µm filtered) | Conventional 2X GMEM (0.22 µm filtered) |
|---|---|---|---|
| Formulation | 2X GMEM High Glucose + 50 mM HEPES; free of NEAA and tryptose phosphate broth | 1X GMEM, often with TPB | Double-strength, for semisolid culture |
| Final filtration pore size | 0.04 µm | 0.22 µm | 0.22 µm |
| Number of filtration stages | 4 | 1 | 1 |
| Mycoplasma barrier filtration | check_circle | cancel | cancel |
| Endotoxin specification | < 0.05 EU/mL | Often not specified at this level | Often not specified at this level |
| USP <788> particulate compliance | check_circle | cancel | cancel |
| Ultrapure Type 1 water (18.2 MΩ·cm) | check_circle | Varies | Varies |
| ISO 13485:2016 manufacturing | check_circle | Varies | Varies |
| Microfluidic channel compatibility | check_circle | cancel | cancel |
| Custom formulation | check_circle | cancel | cancel |
Frequently asked questions
Common questions about using DCP-GMEMH-JN2X for semisolid, 3D, microfluidic and conventional culture.
Supporting literature
Peer-reviewed background on GMEM and BHK-21 cells, soft-agar and stem-cell culture, HEPES buffering, Phenol Red, organ-on-a-chip culture and mycoplasma control.
- Macpherson I, Stoker M. Polyoma transformation of hamster cell clones—an investigation of genetic factors affecting cell competence. Virology. 1962;16(2):147–151. doi:10.1016/0042-6822(62)90290-8
- Eagle H. Nutrition needs of mammalian cells in tissue culture. Science. 1955;122(3168):501–504. doi:10.1126/science.122.3168.501
- Stoker M, Macpherson I. Syrian hamster fibroblast cell line BHK21 and its derivatives. Nature. 1964;203:1355–1357. doi:10.1038/2031355a0
- Borowicz S, Van Scoyk M, Avasarala S, et al. The soft agar colony formation assay. J Vis Exp. 2014;(92):e51998. doi:10.3791/51998
- Ying QL, Nichols J, Chambers I, Smith A. BMP induction of Id proteins suppresses differentiation and sustains embryonic stem cell self-renewal in collaboration with STAT3. Cell. 2003;115(3):281–292. doi:10.1016/S0092-8674(03)00847-X
- Good NE, Winget GD, Winter W, Connolly TN, Izawa S, Singh RMM. Hydrogen ion buffers for biological research. Biochemistry. 1966;5(2):467–477. doi:10.1021/bi00866a011
- Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci USA. 1986;83(8):2496–2500. doi:10.1073/pnas.83.8.2496
- Bhatia SN, Ingber DE. Microfluidic organs-on-chips. Nat Biotechnol. 2014;32(8):760–772. doi:10.1038/nbt.2989
- Low LA, Mummery C, Berridge BR, Austin CP, Tagle DA. Organs-on-chips: into the next decade. Nat Rev Drug Discov. 2021;20(5):345–361. doi:10.1038/s41573-020-0079-3
- Leung CM, de Haan P, Ronaldson-Bouchard K, et al. A guide to the organ-on-a-chip. Nat Rev Methods Primers. 2022;2:33. doi:10.1038/s43586-022-00118-6
- Drexler HG, Uphoff CC. Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention. Cytotechnology. 2002;39(2):75–90. doi:10.1023/A:1022913015916
- Halldorsson S, Lucumi E, Gómez-Sjöberg R, Fleming RMT. Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices. Biosens Bioelectron. 2015;63:218–231. doi:10.1016/j.bios.2014.07.029