FluxMPS™ Ham's F-10 Nutrient Mixture with 50mM HEPES w/o Sodium Bicarbonate: 2X Liquid
FluxMPS™ Ham's F-10 Nutrient Mixture with 50mM HEPES w/o Sodium Bicarbonate: 2X Liquid
Double-strength (2X) Ham's F-10 with 50 mM HEPES — 2.2 g/L glucose and 50 mM HEPES in the concentrate, 1.1 g/L and 25 mM at 1X — for CHO and clonal culture, established and selected primary cells, cell handling and semisolid culture development, ultra-filtered through a quadruple-stage 0.1 µm / 0.04 µm filtration system. With a 50 mM HEPES buffer, bicarbonate-free, it is approximately 5× cleaner than conventional 0.22 µm-filtered media by particulate count.
- 2X concentrate: mix with an equal volume of a compatible solution for a final 1X medium
- Ham's F-10 nutrients including trace elements, hypoxanthine, thymidine, lipoic acid and vitamin B12
- 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
- 50 mM HEPES (25 mM at 1X) for CO₂-independent pH stability during cell preparation, sampling and microscopy
- Endotoxin < 0.05 EU/mL (USP <85> BET)
- Formulation: [+] 2.2 g/L Glucose, [+] L-Glutamine, [+] Sodium Pyruvate, [+] Phenol Red, [+] 50mM HEPES; [−] Sodium Bicarbonate
- 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-Glucose2,200 mg/L
- L-Glutamine292 mg/L
- Sodium Pyruvate220 mg/L
- Buffer50 mM HEPES (no NaHCO₃)
- Phenol Red2.6 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 Ham's F-10 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™ Ham's F-10 keeps Ham's 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 2,200 mg/L D-glucose, 292 mg/L L-glutamine and 220 mg/L 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
46 components at double strength, so agar, supplements, treatment solutions or cell suspensions 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-H10H-B2X is membrane filtered at 0.1 µm twice and 0.04 µm twice — a ready-to-dilute 2X Ham's F-10 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.
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03
0.1 µmSterile-filtration I
Second-pass redundancy that catches anything shed or bypassed upstream.
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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™ Ham's F-10 is approximately 5× cleaner than conventional 0.22 µm-filtered media, lowering clog risk in narrow channels and long perfusion runs.
© Diagnocine® — DCP-H10H-B2X
Where FluxMPS™ Ham's F-10 fits
Ham’s F-10 Nutrient Mixture With HEPES is a basal medium providing amino acids, vitamins, inorganic salts, glucose, trace elements, and nucleotide precursors for in vitro mammalian-cell research. Originally developed for clonal growth of Chinese hamster ovary (CHO) cells, F-10 is also used with other established cell lines and selected primary-cell cultures when appropriately supplemented.[1,3] The included HEPES buffer increases the medium’s resistance to pH changes, particularly during handling outside a controlled carbon dioxide atmosphere.[6] This makes it useful for validated research workflows involving cell preparation, repeated sampling, and microscope-based observation.
Ham’s F-10 is a basal medium rather than a universally complete growth medium. Serum, growth factors, or other supplements should be selected for the cell type and experimental objective; its history of serum-free CHO culture does not mean it supports every cell type without supplementation.[5]
Ham’s F-10 Nutrient Mixture With HEPES, 2X, is a double-strength version of the corresponding 1X formulation, intended for laboratory preparations that require dilution with an equal volume of a compatible solution. Its purpose is to maintain the intended final basal-medium concentrations after mixing, rather than to expose cells routinely to twice the normal nutrient and salt concentrations. A full 2X formulation also contains twice the intended final HEPES concentration (50 mM in the 2X, 25 mM at 1X) if the second solution contributes no HEPES. All basal nutrients, salts, buffer components, and supplements contributed by either solution must be accounted for in the final formulation.
The dilution-compensation principle is used in soft-agar assays, where equal volumes of 2X medium and aqueous agar produce a final 1X medium.[4] The cited protocol uses DMEM/F-12, not Ham’s F-10, so it supports the principle but does not establish performance of an F-10/HEPES 2X product.
The F-10 nutrients and the HEPES buffer serve different functions: F-10 supplies the basal nutrients, while HEPES provides additional buffering. HEPES is especially useful during work outside the incubator where pH may otherwise drift. The 2X format serves a separate purpose: it compensates for substantial dilution during preparation. A routine culture already using correctly supplemented 1X medium does not need a 2X product simply because it contains HEPES.
R&D Applications:
- CHO cell growth and clonal-culture research
- Established mammalian-cell research
- Selected primary-cell research
- Cell-handling and microscopy studies
- Research-medium optimization
- Buffering-condition comparisons
- Semisolid or matrix-containing culture development
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.
CHO Growth & Clonal Culture
Ham's F-10 was developed for clonal growth of Chinese hamster ovary cells and remains a reference basal medium for CHO research.[1,3]
Established & Primary Cells
Used with other established lines and selected primary-cell cultures when appropriately supplemented with serum or growth factors.[1]
Cell Handling & Microscopy
25 mM HEPES at 1X resists pH drift during cell preparation, repeated sampling and microscope-based observation.[6]
Semisolid & Matrix Culture
The 2X format compensates for dilution when developing soft-agar or matrix-containing preparations and optimizing research media.[4]
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-H10H-B2X. Lot-specific values are reported on the Certificate of Analysis.
| Parameter | Specification |
|---|---|
| Formulation | [+] 2.2 g/L Glucose, [+] L-Glutamine, [+] Sodium Pyruvate, [+] Phenol Red, [+] 50mM HEPES; [−] Sodium Bicarbonate |
| 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 | 2,200 mg/L (2.2 g/L; 1.1 g/L at 1X) |
| L-Glutamine | 292 mg/L |
| Sodium Pyruvate | 220 mg/L |
| Sodium Bicarbonate | Not included |
| Phenol Red | 2.6 mg/L |
| HEPES | 11,916 mg/L (50 mM; 25 mM at 1X) |
| 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 | Not required for HEPES buffering (50 mM in the 2X, 25 mM at 1X); add sodium bicarbonate if the final medium will be held in a CO₂ incubator |
| 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-H10H-B2X is 2X Ham's F-10 with 2.2 g/L glucose, L-glutamine, sodium pyruvate, Phenol Red and 50 mM HEPES. It does not contain sodium bicarbonate. Users are advised to review the literature for recommendations regarding medium supplementation and physiological growth requirements specific for different cell lines. All 46 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 | 88.200 |
| Copper sulphate pentahydrate | 7758-99-8 | 0.0050 |
| Ferrous sulphate heptahydrate | 7782-63-0 | 1.668 |
| Magnesium sulphate anhydrous | 7487-88-9 | 149.280 |
| Potassium chloride | 7447-40-7 | 570.000 |
| Potassium phosphate monobasic | 7778-77-0 | 166.000 |
| Sodium chloride | 7647-14-5 | 14800.000 |
| Sodium phosphate dibasic anhydrous | 7558-79-4 | 307.400 |
| Zinc sulphate heptahydrate | 7446-20-0 | 0.0576 |
| Component | CAS Number | mg/L |
|---|---|---|
| AMINO ACIDS | ||
| Glycine | 56-40-6 | 15.020 |
| L-Alanine | 56-41-7 | 17.820 |
| L-Arginine hydrochloride | 1119-34-2 | 422.000 |
| L-Asparagine monohydrate | 5794-13-8 | 30.020 |
| L-Aspartic acid | 56-84-8 | 26.600 |
| L-Cysteine hydrochloride monohydrate | 7048-04-6 | 70.260 |
| L-Glutamic acid | 56-86-0 | 29.400 |
| L-Glutamine | 56-85-9 | 292.000 |
| L-Histidine hydrochloride monohydrate | 5934-29-2 | 42.000 |
| L-Isoleucine | 73-32-5 | 5.200 |
| L-Leucine | 61-90-5 | 26.200 |
| L-Lysine hydrochloride | 657-27-2 | 58.600 |
| L-Methionine | 63-68-3 | 8.960 |
| L-Phenylalanine | 63-91-2 | 9.920 |
| L-Proline | 147-85-3 | 23.000 |
| L-Serine | 56-45-1 | 21.000 |
| L-Threonine | 72-19-5 | 7.140 |
| L-Tryptophan | 73-22-3 | 1.200 |
| L-Tyrosine disodium salt dihydrate | 122666-87-4 | 5.220 |
| L-Valine | 72-18-4 | 7.000 |
| Component | CAS Number | mg/L |
|---|---|---|
| VITAMINS | ||
| Biotin | 58-85-5 | 0.048 |
| Choline chloride | 67-48-1 | 1.396 |
| D-Ca-Pantothenate | 137-08-6 | 1.430 |
| Folic acid | 59-30-3 | 2.640 |
| Nicotinamide | 98-92-0 | 1.230 |
| Pyridoxine hydrochloride | 58-56-0 | 0.412 |
| Riboflavin | 83-88-5 | 0.752 |
| Thiamine hydrochloride | 67-03-8 | 2.000 |
| Vitamin B12 | 68-19-9 | 2.720 |
| i-Inositol | 87-89-8 | 1.082 |
| OTHERS | ||
| D-Glucose | 50-99-7 | 2200.000 |
| HEPES | 7365-45-9 | 11916.000 |
| Hypoxanthine sodium salt | 45738-97-4 | 8.160 |
| Lipoic acid | 1077-28-7 | 0.420 |
| Phenol Red | 34487-61-1 | 2.600 |
| Sodium pyruvate | 113-24-6 | 220.000 |
| Thymidine | 50-89-5 | 1.460 |
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-H10H-B2X compares
Same Ham's F-10 chemistry, different purity class. Conventional columns describe typical single-pass 0.22 µm-filtered catalog media.
| Parameter | DCP-H10H-B2X (FluxMPS™) | Conventional Ham's F-10, 1X (0.22 µm filtered) | Conventional 2X Ham's F-10 (0.22 µm filtered) |
|---|---|---|---|
| Formulation | 2X Ham's F-10 + 50 mM HEPES; free of sodium bicarbonate | 1X F-10, ready to use | 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-H10H-B2X for semisolid, 3D, microfluidic and conventional culture.
Supporting literature
Peer-reviewed background on Ham's F-10 and CHO culture, HEPES buffering, soft-agar culture, Phenol Red, organ-on-a-chip culture and mycoplasma control.
- Ham RG. An improved nutrient solution for diploid Chinese hamster and human cell lines. Exp Cell Res. 1963;29(3):515–526.
- Eagle H. Nutrition needs of mammalian cells in tissue culture. Science. 1955;122(3168):501–504. doi:10.1126/science.122.3168.501
- Tjio JH, Puck TT. Genetics of somatic mammalian cells. II. Chromosomal constitution of cells in tissue culture. J Exp Med. 1958;108(2):259–268. doi:10.1084/jem.108.2.259
- 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
- Ham RG. Clonal growth of mammalian cells in a chemically defined, synthetic medium. Proc Natl Acad Sci USA. 1965;53(2):288–293. doi:10.1073/pnas.53.2.288
- 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
