FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification
Select See the table below
- See the table below
FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification
Ham's F-12 nutrient mixture is a widely utilized basal medium for culturing a variety of mammalian cells, including Chinese Hamster Ovary (CHO) cells, cancer cells, primary cells, and chicken embryonic cells. Developed by R.G. Ham in 1965 as an enhancement over the earlier F-10 nutrient mixture, it contains a more comprehensive array of components such as zinc, putrescine, hypoxanthine, and thymidine compared to other basal media like DMEM. Formulated for serum-free culture of single cells and clonal growth, Ham's F-12 can also be supplemented with serum for other cell types. While lacking proteins, lipids, and growth factors, it requires supplementation with serum or serum-free additives to facilitate optimal growth. The medium employs a sodium bicarbonate buffer system, necessitating a 5-10% CO2 environment to maintain physiological pH.
Ham's F-12 nutrient mixture, Coon's Modification, is a variant of the original Ham's F-12 medium developed specifically for culturing mammalian cells. This modified version was created to determine optimal conditions for obtaining viable hybrids by treating parental cells with Sendai virus. The modification involves doubling the concentrations of amino acids and sodium pyruvate compared to the original F-12 formulation, as well as the addition of ascorbic acid (vitamin C), which was not present in the original mixture. The salt concentrations have also been altered in this modified version. This modification is specifically tailored for hybridoma production and culturing certain rodent cell lines, with increased amino acid and vitamin concentrations, aiming to provide an optimized environment for cell growth and hybridization processes in mammalian cell culture applications. Every FluxMPS™ variant is purified through quadruple-stage 0.04 micron filtration, engineered for organ-on-chip (OoC), tissue-on-chip (ToC), and lab-on-chip (LoC) microfluidic culture systems.
- 4 variants across Sodium Bicarbonate inclusion/exclusion and standard (2.0 g/L) or High Glucose (L-Glutamine, Pyruvate, and Phenol Red included in every variant)
- Doubled amino acid and sodium pyruvate concentrations relative to the original Ham's F-12 formulation
- Ascorbic acid (vitamin C) added — not present in the original F-12 mixture
- Altered salt concentrations relative to the original F-12 formulation
- Specifically tailored for hybridoma production via Sendai virus cell fusion and culturing certain rodent cell lines
- Sodium bicarbonate buffering system requiring a 5-10% CO2 environment
- FluxMPS™ quadruple-stage 0.04 micron filtration — finer than any ready-to-use cell culture media currently available
- Available in 500 mL and 1000 mL sizes; 2-8°C storage away from bright light; fully customizable on request (including HEPES)
- Concentration1X
- Glucose2.0 g/L standard, or High Glucose
- L-Glutamine / PyruvateIncluded (all variants)
- Sodium BicarbonateConfigurable (+/-)
- Phenol RedIncluded (all variants)
- Ascorbic Acid (Vitamin C)Added vs. original F-12
- Sizes500 mL / 1000 mL
- Storage2-8 C, away from light
Select Your Ham's F-12 Coon's Modification Configuration
Please select the supplement(s) of interest, then click Search. Check the supplement(s) you need below and press Search to instantly highlight every matching variant. Each row links straight through to its product page via the catalog number or the View button.
| Name | Cat No. | Glucose | L-Glutamine | Pyruvate | Bicarbonate | Phenol Red | Product Page |
|---|---|---|---|---|---|---|---|
| F12 / Coon's | DCP-F12HCM1X | 2.0 g/L | check | check | check | check | Viewarrow_forward |
| F12 / Coon's w/o Bicarbonate | DCP-12C-B1X | 2.0 g/L | check | check | remove | check | Viewarrow_forward |
| F12 / Coon's High Glucose | DCP-12CHG1X | High | check | check | check | check | Viewarrow_forward |
| F12 / Coon's High Glucose w/o Bicarbonate | DCP-12CHG-B1X | High | check | check | remove | check | Viewarrow_forward |
Reference (Peer-Reviewed Publication)
This product was cited in a peer-reviewed research article:
"Infection dynamics and virulence potential of clinical Pseudomonas aeruginosa isolates in a human airway epithelium model system."
In this study, the researchers used Coon's Modified Ham's F-12 Medium (Diagnocine) as described in the publication.
Corresponding authors:
Claudia Antonella Colque (antcol@biosustain.dtu.dk)
Ruggero La Rosa (rugros@dtu.dk)
Helle Krogh Johansen (hkj@biosustain.dtu.dk)
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark (DTU)
About Ham's F-12 Nutrient Mixture, Coon's Modification
Ham's F-10 and Ham's F-12 are chemically defined, nutrient-rich cell culture media developed by Richard G. Ham at the University of Colorado to support the clonal growth of single Chinese hamster ovary (CHO) cells, an objective that required sustaining individual cells at very low densities under defined conditions, without reliance on serum or undefined biological extracts. They belong to a larger family of sequentially numbered formulations (the "F" series) developed by Ham in the late 1950s-1960s, each iteration tested against clonal growth efficiency as the quantitative readout.
F-12 is a direct development of F-10, retaining the same qualitative amino acid and vitamin profiles but with substantially altered concentrations of several components, most notably an approximately 30-fold higher zinc sulfate level, along with the addition of putrescine and linoleic acid, thereby rendering it effective for low-density, protein-free CHO culture. Ham, R.G. (1965). "Clonal growth of mammalian cells in a chemically defined, synthetic medium." Proceedings of the National Academy of Sciences USA, 53(2): 288-293. PMID: 14283412. F-12 became a medium of choice for myeloma and hybridoma cloning, primary rat hepatocytes, rat prostate epithelial cells, clonal toxicity assays, and the base of the widely used DMEM/F-12 hybrid.
Coon's Modification
Ham's F-12 nutrient mixture, Coon's Modification, is a variant of the original Ham's F-12 medium developed specifically for culturing mammalian cells. This modified version was created to determine optimal conditions for obtaining viable hybrids by treating parental cells with Sendai virus. The modification involves doubling the concentrations of amino acids and sodium pyruvate compared to the original F-12 formulation, as well as the addition of ascorbic acid (vitamin C), which was not present in the original mixture. The salt concentrations have also been altered in this modified version.
This modification is specifically tailored for hybridoma production and culturing certain rodent cell lines, with increased amino acid and vitamin concentrations. This modification aims to provide an optimized environment for cell growth and hybridization processes in mammalian cell culture applications. Coon's modification of F-12 was developed for virally fused hybrid cells — a medium of choice for cloning, widely used in monoclonal antibody work.
Richard G. Ham and the Serum-Free Cloning Problem
By the late 1950s, growing a single mammalian cell into a clone essential for genetic analysis, mutagenesis, and selection of stable lines still depended on high serum concentrations or feeder cells. Ham recognized that the barrier was not a single missing factor but an incompletely defined nutrient environment whose inadequacies were masked by the serum's complex activity.
Working with near-diploid Chinese hamster (CHO) cells, the line initiated by Theodore T. Puck at the University of Colorado in 1957, Ham systematically developed a series of nutrient mixtures using single-cell plating efficiency (the fraction of individually plated cells that formed visible colonies) as his quantitative endpoint. Ham's F-10 (1963) preceded F-12: Ham, R.G. (1963). "An improved nutrient solution for diploid Chinese hamster and human cell lines." Experimental Cell Research, 29: 515-526. PMID: 13952250. F-10 enabled consistent single-cell cloning of Chinese hamster cells under low-serum conditions and was one of the first defined media to incorporate copper (CuSO4) and zinc (ZnSO4) as trace elements, along with hypoxanthine, thymidine, and lipoic acid.
In the 1965 PNAS paper, Ham demonstrated for the first time that single CHO cells could proliferate into clones in a fully chemically defined, protein-free medium. The key changes from F-10 to F-12 were: zinc sulfate increased approximately 30-fold (0.029 to 0.863 mg/L); addition of putrescine * 2HCl (0.161 mg/L), a polyamine precursor; addition of linoleic acid (0.084 mg/L), an essential omega-6 fatty acid; large increases in several amino acids (proline, threonine, tryptophan, tyrosine, valine, isoleucine, lysine); increases in choline chloride and myo-inositol; and a magnesium source change from MgSO4 to MgCl2.
Ham's F-12 Base Composition
The values below reflect the standard Ham's F-12 nutrient mixture (with L-glutamine; without sodium bicarbonate) that Coon's Modification is based upon. All concentrations in mg/L. Coon's Modification doubles the amino acid and sodium pyruvate concentrations shown below, adds ascorbic acid (vitamin C, not present in the original mixture), and alters the salt concentrations; exact per-component values for these Coon's-specific changes are available on request via support@diagnocine.com.
Inorganic Salts (Standard F-12 Base)
| Salt | mg/L |
|---|---|
| Calcium chloride dihydrate (CaCl2 * 2H2O) | 44.100 |
| Copper sulfate pentahydrate (CuSO4 * 5H2O) | 0.0025 |
| Ferrous sulfate heptahydrate (FeSO4 * 7H2O) | 0.834 |
| Magnesium chloride anhydrous (MgCl2) | 57.650 |
| Potassium chloride (KCl) | 223.600 |
| Sodium chloride (NaCl) | 7599.000 |
| Sodium phosphate dibasic anhydrous (Na2HPO4) | 142.040 |
| Zinc sulfate heptahydrate (ZnSO4 * 7H2O) | 0.863 |
Amino Acids (Standard F-12 Base, Before Coon's Doubling)
| Amino Acid | mg/L |
|---|---|
| Glycine | 7.500 |
| L-Alanine | 8.910 |
| L-Arginine hydrochloride | 210.700 |
| L-Asparagine monohydrate | 15.010 |
| L-Aspartic acid | 13.300 |
| L-Cysteine hydrochloride monohydrate | 35.120 |
| L-Glutamic acid | 14.700 |
| L-Glutamine | 146.000 |
| L-Histidine hydrochloride monohydrate | 20.960 |
| L-Isoleucine | 3.940 |
| L-Leucine | 13.100 |
| L-Lysine hydrochloride | 36.500 |
| L-Methionine | 4.480 |
| L-Phenylalanine | 4.960 |
| L-Proline | 34.500 |
| L-Serine | 10.500 |
| L-Threonine | 11.900 |
| L-Tryptophan | 2.040 |
| L-Tyrosine disodium salt dihydrate | 7.810 |
| L-Valine | 11.700 |
Vitamins (Standard F-12 Base)
| Vitamin | mg/L |
|---|---|
| Biotin | 0.0073 |
| Choline chloride | 13.960 |
| D-Ca-Pantothenate | 0.480 |
| Folic acid | 1.320 |
| Nicotinamide | 0.037 |
| Pyridoxine hydrochloride | 0.062 |
| Riboflavin | 0.038 |
| Thiamine hydrochloride | 0.340 |
| Vitamin B12 | 1.360 |
| myo-Inositol | 18.000 |
Other Components (Standard F-12 Base)
| Component | mg/L |
|---|---|
| D-Glucose | 1801.600 |
| Hypoxanthine sodium salt | 4.770 |
| Linoleic acid | 0.084 |
| Lipoic acid | 0.210 |
| Phenol red sodium salt | 1.240 |
| Putrescine dihydrochloride | 0.161 |
| Sodium pyruvate | 110.100 |
| Thymidine | 0.730 |
| Sodium bicarbonate (NaHCO3) | 1176.000 |
Validated Cell Types and Applications
Ham's F-10
CHO / Chinese Hamster Cells
Founding application; clonal growth at low serum (serum-free with albumin/fetuin).
Human Diploid Cells
Fibroblasts and related primary cells at low serum.
White Blood Cells
Chromosome analysis from primary blood cultures.
Primary Rat, Rabbit & Chicken Explants
Supported across multiple species-derived primary tissue explants.
Chick Embryo Cells
Serum-free monolayer culture.
Mouse L Cells & HeLa Cells
Mouse L cells included in the original F-10 cell-type survey; HeLa cells in serum-supplemented culture. Mammalian embryo culture (bovine, equine, porcine, and human embryo systems) has also used Ham's F-10, often F-10 + HEPES, as a holding/culture medium.
Ham's F-12 (Reference)
F-12 is the direct descendant of F-10 and is validated for CHO cells (primary application; protein-free clonal growth; serum-free bioproduction standard, using CHO cells from the Puck line, including the widely used proline-requiring CHO-K1 subclone), mouse L cells (serum-supplemented maintenance), HeLa cells (serum-free culture with defined hormones: insulin, transferrin, hydrocortisone, EGF, FGF), myeloma and hybridoma cells (a medium of choice for cloning, widely used in monoclonal antibody work; Coon's modification of F-12 was developed for virally fused hybrid cells), primary rat hepatocytes, rat prostate epithelial cells, normal rat kidney (NRK) cells, and clonal toxicity assays.
Shared role: base for defined media development. F-12 is the base for the DMEM/F-12 1:1 hybrid (combining DMEM's high amino acid and glucose content with F-12's trace elements, lipids, and low osmolality), one of the most widely used media in serum-free and stem cell culture. It is also the starting formulation from which Ham and colleagues derived the MCDB series for specific primary cell types.
Media Lineage Comparison
| Feature | Ham's F-10 | Ham's F-12 | DMEM (High Glucose) |
|---|---|---|---|
| Developer & year | R.G. Ham, 1963 | R.G. Ham, 1965 | Dulbecco & Freeman, 1959 |
| Parent formulation | Earlier "F" series | F-10 (direct) | BME |
| Trace metals (Cu, Zn, Fe) | Cu + Zn + Fe | Cu + Zn + Fe | Fe only (ferric nitrate) |
| Hypoxanthine | Yes, 4.08 mg/L | Yes, 4.77 mg/L | No |
| Thymidine | Yes, 0.73 mg/L | Yes, 0.73 mg/L | No |
| Lipoic acid | Yes, 0.21 mg/L | Yes, 0.21 mg/L | No |
| Putrescine | No | Yes, 0.161 mg/L | No |
| Linoleic acid | No | Yes, 0.084 mg/L | No |
| Biotin | Yes, 0.024 mg/L | Yes, 0.0073 mg/L | No |
| Vitamin B12 | Yes, 1.36 mg/L | Yes, 1.36 mg/L | No |
| Vitamin B6 form | Pyridoxine HCl | Pyridoxine HCl | Pyridoxine HCl (Gibco) |
| myo-Inositol | 0.541 mg/L | 18.0 mg/L | Present (lower) |
| Glucose | 1100 mg/L (approximately 6.1 mM) | 1801.6 mg/L (approximately 10 mM) | 4500 mg/L (25 mM) |
| Sodium pyruvate | 110 mg/L | 110 mg/L | 110 mg/L (optional) |
| NaHCO3 | 1200 mg/L | 1176 mg/L | 3700 mg/L |
| Serum-free design | Partial (with proteins) | Full (protein-free CHO) | No |
| Osmolality (with NaHCO3) | 300-340 mOsm/kg | 260-300 mOsm/kg | approximately 320-355 mOsm/kg |
Key Structural Differences: F-10 to F-12
Context on the biochemical rationale behind the F-10-to-F-12 evolution that underlies the Coon's Modification base formulation.
The zinc difference is the defining feature. The approximately 30-fold increase in ZnSO4 * 7H2O from F-10 (0.029) to F-12 (0.863 mg/L) was the single most important change enabling protein-free CHO cloning. Zinc is required for numerous metalloenzymes, receptors, and transcription factors; at very low cell densities, the per-volume zinc requirement is not met by trace contamination, so it must be supplied at a defined, elevated concentration.
Putrescine supports polyamine-dependent proliferation. Putrescine (0.161 mg/L) is the diamine precursor to spermidine and spermine, essential for DNA synthesis, ribosome function, and cell division, partially bypassing the polyamine supply that serum normally provides.
Linoleic acid is an essential fatty acid. Linoleic acid (0.084 mg/L, free fatty acid) supplies the essential omega-6 fatty acid for membrane phospholipid synthesis that serum albumin-bound fatty acids would otherwise provide; cells cannot synthesize it de novo.
myo-Inositol is 33x higher in F-12 (18.0 vs 0.541 mg/L). Inositol is a component of phosphatidylinositol, a key membrane phospholipid and signaling intermediate whose requirement rises sharply in single-cell cultures.
Choline is 20x higher in F-12 (13.96 vs 0.698 mg/L), supporting phosphatidylcholine synthesis and the higher membrane biosynthetic demand of rapidly proliferating single-cell clones under protein-free conditions.
Why FluxMPS™ Ham's F-12 Coon's Modification
Quadruple-Stage 0.04 Micron Filtration
Purified to 0.04 microns — finer than any ready-to-use cell culture media currently available — removing the microscopic particulates and protein aggregates that silently block micro-channels.
Built for OoC / ToC / LoC Platforms
Engineered from the ground up for Organ-on-Chip, Tissue-on-Chip, and Lab-on-Chip platforms, where the medium itself is part of the instrument and cannot be an afterthought.
Particulate & Aggregate Removal
Eliminates particulates and protein aggregates that disrupt laminar flow and generate false biological signals before the media ever reaches the chip.
Optical Clarity
Supports real-time imaging and integrated biosensing on Organ-on-Chip platforms, where optical clarity of the medium is essential to data quality.
FDA-Recognized Physiological Modeling Standards
Formulated to support FDA-recognized physiological modeling standards, providing a validated, reproducible media foundation for regulatory-facing studies.
Drug Discovery, Toxicology & Translational Research
A validated foundation for drug discovery, toxicology screening, and translational research, where downstream data must stand up to regulatory scrutiny.
Quadruple-Stage Filtration System
Every FluxMPS™ Ham's F-12 Coon's Modification variant passes through the same four-stage architecture before reaching your chip.
-
01
Pre-Filtration Stage 1 0.1 µm
Initial coarse particulate removal.
-
02
Pre-Filtration Stage 2 0.1 µm
Secondary particulate and aggregate reduction.
-
03
Sterile Filtration Stage 1 0.04 µm
Fine sterile filtration below standard 0.22 micron practice.
-
04
Sterile Filtration Stage 2 0.04 µm
Final polish for microfluidic-grade clarity.
Engineered for Flow, Not Just Growth
Every component is optimized for consistent, laminar flow performance across complex micro-channel geometries, capillary-bed and vascular simulations, and long-term automated perfusion studies running continuously for weeks.
Frequently Asked Questions
Verified Bibliography
- Ham, R.G. (1963). An improved nutrient solution for diploid Chinese hamster and human cell lines. Experimental Cell Research, 29: 515-526. PMID: 13952250. DOI: 10.1016/S0014-4827(63)80014-2
- Ham, R.G. (1965). Clonal growth of mammalian cells in a chemically defined, synthetic medium. Proceedings of the National Academy of Sciences USA, 53(2): 288-293. PMID: 14283412. DOI: 10.1073/pnas.53.2.288
- Ham, R.G. & McKeehan, W.L. (1979). Media and growth requirements. Methods in Enzymology, 58: 44-93. DOI: 10.1016/S0076-6879(79)58127-2
- Barnes, D. & Sato, G. (1980). Serum-free cell culture: a unifying approach. Cell, 22(3): 649-655. DOI: 10.1016/0092-8674(80)90540-1
- McKeehan, W.L., Hamilton, W.G. & Ham, R.G. (1976). Selenium is an essential trace nutrient for the growth of WI-38 diploid human fibroblasts. Proc. Natl. Acad. Sci. USA, 73(6): 2023-2027. DOI: 10.1073/pnas.73.6.2023
- Yao, T. & Asayama, Y. (2017). Animal-cell culture media: History, characteristics, and current issues. Reproductive Medicine and Biology, 16(2): 99-117. PMC5661806
FluxMPS™ — Precision Cell Culture Media for Microphysiological Systems
Built for the architecture of the future. Not the flask of the past.
Traditional cell culture media were formulated for static well plates and flasks — environments that tolerate impurities, precipitates, and particle loads that would immediately compromise a microfluidic system. FluxMPS™ was designed from the ground up for Organ-on-Chip (OoC), Tissue-on-Chip (ToC), and Lab-on-Chip (LoC) platforms, where the medium itself is part of the instrument.
Purity That Protects Your Platform
FluxMPS™ is purified to 0.04 microns — finer than any ready-to-use cell culture media currently available. At this level, the microscopic particulates and protein aggregates that silently block micro-channels, disrupt laminar flow, and generate false biological signals are eliminated before the media ever reaches your chip.
The result: your platform stays operational, your data stays clean, and your biology drives the result — not your media.
Engineered for Flow, Not Just Growth
The name FluxMPS™ reflects its core design principle. Every component is optimized for consistent, laminar flow performance across:
- Complex micro-channel geometries
- Capillary-bed and vascular simulations
- Long-term automated perfusion studies running continuously for weeks
Zero-clogging performance is not a feature — it is the baseline specification.
Applications & Performance
| Application | What FluxMPS™ Delivers |
|---|---|
| Microfluidics | Stable shear stress; no channel blockage |
| Metabolic Tracing | Ultra-pure matrix with no contaminant interference |
| Long-term Perfusion | Consistent formulation stability over weeks of continuous flow |
| Organ-on-Chip | Optical clarity for real-time imaging and integrated biosensing |
Regulatory Foundation
FluxMPS™ is formulated to support FDA-recognized physiological modeling standards, providing a validated, reproducible media foundation for drug discovery, toxicology screening, and translational research. When your downstream data needs to stand up to regulatory scrutiny, your upstream media cannot be an afterthought.
The Bottom Line
Microfluidic platforms are precision instruments. They require precision inputs.
FluxMPS™ is the only ready-to-use cell culture medium engineered specifically to meet that standard — protecting your chip, your cells, and your science.
