FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification [2X] Family

verified Double-Strength · Developed by R.G. Ham, 1965; Coon's Modification for Hybridoma Production

FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification [2X] Family

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. This [2X] family supplies the Coon's formulation at double strength — 4.0 g/L glucose in the concentrate, 2.0 g/L at 1X (High Glucose: 9.0 g/L and 4.5 g/L) — for preparations where agar, agarose, or another solution would otherwise dilute the basal medium. 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 — DCP-F12HCM2X, DCP-12C-B2X, DCP-12CHG2X, DCP-12CHG-B2X — across Sodium Bicarbonate inclusion/exclusion and standard (4.0 g/L at 2X) or High Glucose (9.0 g/L at 2X) (L-Glutamine, Pyruvate, and Phenol Red included in every variant; no variant contains HEPES)
2X concentrate: mix with an equal volume of a compatible solution or matrix for a final 1X medium
Doubled amino acid and sodium pyruvate concentrations relative to the original Ham's F-12 formulation — at 2X: 584 mg/L L-glutamine and 440 mg/L sodium pyruvate; sodium bicarbonate 5,352 mg/L where included
Ascorbic acid (vitamin C) added — not present in the original F-12 mixture (90 mg/L at 2X)
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; the 2X product pages also list FRTL-5 and other rat thyroid cells, hormone-response and iodide-transport research, and agarose culture
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 a 500 mL size; 2-8°C storage away from bright light, do not freeze; fully customizable on request (including HEPES)

SKU: HamsF-12NutrientMixtureCoonsModification2X — 4 VARIANTS

FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification — 2X Liquid Cell Culture Medium

Concentration2X (double strength)
Working strength1X after 1:1 mixing
Glucose4.0 g/L at 2X (2.0 g/L at 1X), or High Glucose 9.0 g/L (4.5 g/L at 1X)
L-Glutamine / PyruvateIncluded (all variants)
Sodium BicarbonateConfigurable (+/-)
Phenol RedIncluded (all variants)
Ascorbic Acid (Vitamin C)Added vs. original F-12
FiltrationQuadruple-stage, 0.04 µm final
Size500 mL
Storage2-8 C, away from light; do not freeze

RUOHam 1965 / Coon's Modification2X ConcentrateCustomizable

Product Selector

Select Your Ham's F-12 Coon's Modification [2X] 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.

At-a-glance supplement matrix — 4 variants

At-a-glance supplement matrix — click to view product page. L-Glutamine, Pyruvate, and Phenol Red are included in every variant. Glucose is shown at 2X. HEPES buffering is available as a customization on request but is not a standard catalog option.

Name	Cat No.	Glucose	L-Glutamine	Pyruvate	Bicarbonate	Phenol Red	Product Page
F12 / Coon's	DCP-F12HCM2X	4.0 g/L	check	check	check	check	Viewarrow_forward
F12 / Coon's w/o Bicarbonate	DCP-12C-B2X	4.0 g/L	check	check	remove	check	Viewarrow_forward
F12 / Coon's High Glucose	DCP-12CHG2X	9.0 g/L (High)	check	check	check	check	Viewarrow_forward
F12 / Coon's High Glucose w/o Bicarbonate	DCP-12CHG-B2X	9.0 g/L (High)	check	check	remove	check	Viewarrow_forward

check Included remove Not included All variants: 2X concentration · L-Glutamine · Sodium Pyruvate · Phenol Red · Ascorbic Acid · no HEPES · 500 mL

Customization available: All variants above use the standard 2X concentration (mix 1:1 for 1X). Standard Glucose concentration is 4.0 g/L at 2X (2.0 g/L at 1X); a High Glucose option is also available (see matrix above). 25 mM HEPES is available as a standard customization concentration. Other concentrations, additions of chemicals, compounds, proteins, supplements, a different pH, and modifications are available on request — contact support@diagnocine.com.

Cited in Peer-Reviewed Research

Reference (Peer-Reviewed Publication)

Diagnocine's Coon's Modified Ham's F-12 Medium 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)

Download Full PDFarrow_forward View Journal Article (bioRxiv)arrow_forward

About Coon's Modification

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.^[2] 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.^[7] 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. Originally developed for hybrid-cell culture, Coon's modified F-12 is also used, with appropriate supplements, for rat thyroid cell lines such as FRTL-5,^[7,8] providing a basal nutrient environment for laboratory studies of cell growth, hormone responsiveness, and differentiated thyroid-cell functions.

Why a 2X (Double-Strength) Format

Ham's F-12 Nutrient Mixture, Coon's Modification, 2X, is a double-strength version of the corresponding 1X Coon's formulation, intended for laboratory workflows that dilute the basal medium during final preparation. Its purpose is dilution compensation, following the established use of 2X media in equal-volume mixtures with aqueous agar or agarose preparations.^[9] Mixing one volume of 2X medium with one volume of a compatible solution that contributes none of its basal ingredients gives those ingredients at their intended 1X concentrations: D-glucose 4,000 mg/L to 2,000 mg/L (High Glucose 9,000 mg/L to 4,500 mg/L), L-glutamine 584 mg/L to 292 mg/L, sodium pyruvate 440 mg/L to 220 mg/L, and, where included, sodium bicarbonate 5,352 mg/L to 2,676 mg/L. The published agar/agarose methods establish this dilution principle, not the compatibility or performance of a specific 2X Coon's product.

Important distinction: Ordinary 1X Coon's modification already has approximately double the amino acids and pyruvate of standard F-12; that does not make it a 2X medium. “2X Coon's” means twice the corresponding Coon's formulation, not simply another name for standard-strength Coon's modified F-12.

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.^[1] 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.

Composition

Coon's Modification [2X] Composition

Concentration scale: The Coon's Modification tables below list every component at 2X as given on the 2X product pages; the 1X column is each value halved, since the product pages state that values are halved after 1:1 dilution. The complete formulation lists 50 components; the w/o Sodium Bicarbonate variants list 49. The standard Ham's F-12 base tables that follow are given at 1X for reference.

Component Summary

Concentration	2X (double strength); 1X after 1:1 mixing
D-Glucose	4,000 mg/L at 2X (2.0 g/L at 1X); High Glucose 9,000 mg/L at 2X (4.5 g/L at 1X)
L-Glutamine	584 mg/L at 2X (all variants)
Sodium Pyruvate	440 mg/L at 2X (all variants)
Buffer	NaHCO3 5,352 mg/L at 2X where included; no HEPES
Phenol Red	2.48 mg/L at 2X (all variants)
Ascorbic Acid	90 mg/L at 2X (all variants)
Appearance	Red to bright pink

Coon's Modification — Inorganic Salts

Salt	CAS No.	2X as supplied (mg/L)	1X after 1:1 mixing (mg/L)
Calcium chloride dihydrate	10035-04-8	330.000	165.000
Copper sulphate pentahydrate	7758-99-8	0.0050	0.0025
Disodium hydrogen phosphate anhydrous	7558-79-4	264.080	132.040
Ferrous sulphate heptahydrate	7782-63-0	1.668	0.834
Magnesium chloride hexahydrate	7791-18-6	212.000	106.000
Magnesium sulphate anhydrous	7487-88-9	101.600	50.800
Potassium chloride	7447-40-7	610.000	305.000
Potassium dihydrogen phosphate	7778-77-0	136.000	68.000
Sodium bicarbonate (not in the w/o Sodium Bicarbonate variants)	144-55-8	5352.000	2676.000
Sodium chloride	7647-14-5	15060.000	7530.000
Zinc sulphate heptahydrate	7446-20-0	0.288	0.144

Coon's Modification — Amino Acids

Amino Acid	CAS No.	2X as supplied (mg/L)	1X after 1:1 mixing (mg/L)
Glycine	56-40-6	32.000	16.000
L-Alanine	56-41-7	36.000	18.000
L-Arginine hydrochloride	1119-34-2	844.000	422.000
L-Asparagine anhydrous	70-47-3	60.000	30.000
L-Aspartic acid	56-84-8	52.000	26.000
L-Cysteine hydrochloride	52-89-1	140.520	70.260
L-Glutamic acid	56-86-0	60.000	30.000
L-Glutamine	56-85-9	584.000	292.000
L-Histidine hydrochloride monohydrate	5934-29-2	84.000	42.000
L-Isoleucine	73-32-5	15.600	7.800
L-Leucine	61-90-5	52.400	26.200
L-Lysine hydrochloride	657-27-2	146.000	73.000
L-Methionine	63-68-3	18.000	9.000
L-Phenylalanine	63-91-2	20.000	10.000
L-Proline	147-85-3	140.000	70.000
L-Serine	56-45-1	42.000	21.000
L-Threonine	72-19-5	47.600	23.800
L-Tryptophan	73-22-3	8.000	4.000
L-Tyrosine disodium salt	69847-45-6	31.720	15.860
L-Valine	72-18-4	46.800	23.400

Coon's Modification — Vitamins

Vitamin	CAS No.	2X as supplied (mg/L)	1X after 1:1 mixing (mg/L)
Biotin	58-85-5	0.0146	0.0073
Choline chloride	67-48-1	27.920	13.960
D-Ca-Pantothenate	137-08-6	0.960	0.480
Folic acid	59-30-3	2.640	1.320
L-Ascorbic acid (added in Coon's Modification)	50-81-7	90.000	45.000
Nicotinamide	98-92-0	0.080	0.040
Pyridoxine hydrochloride	58-56-0	0.120	0.060
Riboflavin	83-88-5	0.080	0.040
Thiamine hydrochloride	67-03-8	0.680	0.340
Vitamin B12	68-19-9	2.720	1.360
i-Inositol	87-89-8	36.040	18.020

Coon's Modification — Other Components

Component	CAS No.	2X as supplied (mg/L)	1X after 1:1 mixing (mg/L)
D-Glucose (standard)	50-99-7	4000.000	2000.000
D-Glucose (High Glucose variants)	50-99-7	9000.000	4500.000
Hypoxanthine sodium salt	45738-97-4	8.080	4.040
Linoleic acid	60-33-3	0.180	0.090
Lipoic acid	1077-28-7	0.412	0.206
Phenol Red	34487-61-1	2.480	1.240
Putrescine dihydrochloride	333-93-7	0.600	0.300
Sodium pyruvate	113-24-6	440.000	220.000
Thymidine	50-89-5	1.400	0.700

Ham's F-12 Base Composition (Reference, 1X)

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; the per-component Coon's values are listed in the Coon's Modification tables above. Per-lot Certificate of Analysis (CoA) requests can be directed to 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

F-12 uses only dibasic sodium phosphate (Na₂HPO₄), in contrast to F-10's dual KH₂PO₄ + Na₂HPO₄ system. F-10 uses MgSO₄ (anhydrous, 74.64 mg/L); F-12 uses MgCl₂ (anhydrous, 57.65 mg/L), reducing the sulfate load. Coon's Modification further alters these salt concentrations relative to standard F-12.

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

Coon's Modification: doubles the amino acid concentrations shown above, relative to the original F-12 formulation, and also doubles sodium pyruvate. Ascorbic acid (vitamin C) is added, which is not present in the original mixture above.

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

Glucose in this family: the catalog offers a standard formulation (2.0 g/L at 1X; 4.0 g/L at 2X) and a High Glucose formulation (4.5 g/L at 1X; 9.0 g/L at 2X) (see supplement matrix above), both distinct from the standard F-12 base value shown here.

Validated Applications

Validated Cell Types and Applications

Coon's Modification [2X] — Research Applications

The 2X product pages list hybrid-cell culture research, rat thyroid-cell culture, hormone-response and cell-signaling research, iodide-transport and thyroid-function research, cell-based chemical-response studies, chondrogenic differentiation research, and semisolid or agarose culture development.

Endocrinology

FRTL-5 & Rat Thyroid Cells

The basal medium for hormone-dependent FRTL-5 thyroid epithelial cells when supplemented with serum and a defined hormone mixture.^[8]

FRTL-5ThyroidTSH response

Cell Genetics

Hybrid-Cell Culture

Coon's modification was developed for the formation and growth of viable somatic cell hybrids.^[7]

Somatic hybridsCell fusionClonal growth

Physiology

Hormone Response & Iodide Transport

A basal environment for studying hormone responsiveness, cell signaling, iodide uptake, and other differentiated thyroid-cell functions.^[8]

Hormone responseIodide transportSignaling

Differentiation

Chondrogenic & Agarose Culture

A double-strength base for agarose and other semisolid preparations, including chondrogenic differentiation research.^[9]

ChondrogenesisAgaroseSemisolid

Microfluidics

Organ-on-a-Chip & MPS

Microchannel-safe medium for perfused organ-, tissue-, and body-on-a-chip platforms.^[13,14,15]

OoCToCMPS

Live-Cell Imaging

Microscopy & Optical Sensing

Fewer particles mean cleaner fields for colony counting, confocal imaging, and on-chip biosensors.

ConfocalColony countingBiosensors

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.

Comparison

Media Lineage Comparison

F-series lineage behind Coon's Modification; values in this table refer to the standard media at 1X working strength.

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

Ham's F-12 Nutrient Mixture, Coon's Modification: 1X vs. 2X

Feature	Coon's Modification, 1X	Coon's Modification, 2X
Supplied concentration	1X (ready to use)	2X (double strength)
Preparation before use	None	Mix with an equal volume of a compatible solution or matrix (for example aqueous agar or agarose) for a final 1X medium
Component concentrations as supplied	1X Coon's formulation (about twice the amino acids and pyruvate of standard F-12)	Twice the 1X Coon's value (2X column of the composition tables), before dilution
Composition at working strength	Coon's Modification formulation	Coon's Modification formulation, after 1:1 dilution
Glucose	2.0 g/L standard, or High Glucose	4.0 g/L (2.0 g/L after 1:1), or High Glucose 9.0 g/L (4.5 g/L after 1:1)
Sodium bicarbonate (where included)	2,676 mg/L	5,352 mg/L (2,676 mg/L after 1:1 dilution)
Catalog variants	4 (bicarbonate +/-, standard or High Glucose)	4 (bicarbonate +/-, standard or High Glucose)
Catalog number suffix	...1X	...2X (e.g., DCP-F12HCM2X)
Filtration	Quadruple-stage, 0.04 micron final	Quadruple-stage, 0.04 micron final
Sizes	500 mL / 1000 mL	500 mL
Storage	2-8°C, away from bright light	2-8°C, away from bright light; do not freeze

Background

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.^[3,4,5,6]

Why FluxMPS™

Why FluxMPS™ Ham's F-12 Coon's Modification [2X]

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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.

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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.

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Particulate & Aggregate Removal

Eliminates particulates and protein aggregates that disrupt laminar flow and generate false biological signals before the media ever reaches the chip.

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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.

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FDA-Recognized Physiological Modeling Standards

Formulated to support FDA-recognized physiological modeling standards, providing a validated, reproducible media foundation for regulatory-facing studies.

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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.

Filtration Technology

Quadruple-Stage Filtration System

Every FluxMPS™ Ham's F-12 Coon's Modification [2X] variant passes through the same four-stage architecture before reaching your chip: two 0.1 micron pre-filtration stages followed by two 0.04 micron final filtration stages.

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

Final Filtration Stage 1 0.04 µm

Fine filtration below standard 0.22 micron practice.
04

Final 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.

0.04µm

Final polish

4

Total filtration stages

FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification [2X] Family quadruple-stage filtration system diagram ? two 0.1 micron pre-filtration stages followed by two 0.04 micron final filtration stages, engineered for organ-on-a-chip (OoC), tissue-on-a-chip (ToC), and lab-on-a-chip microfluidic cell culture media applications by Diagnocine.

FAQ

Frequently Asked Questions

The family covers combinations of Sodium Bicarbonate inclusion and standard (4.0 g/L at 2X, 2.0 g/L at 1X) or High Glucose (9.0 g/L at 2X, 4.5 g/L at 1X); L-Glutamine, Pyruvate, and Phenol Red are included in every variant, and no variant contains HEPES. Use the supplement matrix above: check the box for each supplement your protocol requires, press Search, and the matching row (or rows) will highlight in mint green so you can click straight through via the catalog number link or the View button.

Mix one volume of 2X Coon's medium with one volume of a compatible solution that contributes none of its basal ingredients — for example aqueous agar or agarose — so glucose (2.0 g/L, or 4.5 g/L for High Glucose), amino acids, vitamins, and salts end up at their 1X Coon's concentration.^[9] Every component falls to half its supplied level: L-glutamine 584 mg/L to 292 mg/L, sodium pyruvate 440 mg/L to 220 mg/L, and sodium bicarbonate 5,352 mg/L to 2,676 mg/L where included. Note that ordinary 1X Coon's already has about double the amino acids and pyruvate of standard F-12; that does not make it a 2X medium. Serum, antibiotics, and other supplements can be added before or after filter sterilization, observing sterility precautions.

Coon's Modification was created to determine optimal conditions for obtaining viable hybrids by treating parental cells with Sendai virus.^[7] It doubles the concentrations of amino acids and sodium pyruvate compared to the original F-12 formulation, adds ascorbic acid (vitamin C, not present in the original mixture), and alters the salt concentrations. It is specifically tailored for hybridoma production and culturing certain rodent cell lines. “2X Coon's” means twice the corresponding Coon's formulation, not simply another name for standard-strength Coon's modified F-12.

Yes. Coon's Modified Ham's F-12 Medium (Diagnocine) was cited in a peer-reviewed study, "Infection dynamics and virulence potential of clinical Pseudomonas aeruginosa isolates in a human airway epithelium model system," by researchers at the Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark (DTU). See the Cited in Peer-Reviewed Research section above for the full citation and links.

F-12 (1965) is a direct development of F-10 (1963), both developed by Richard G. Ham. F-12 increased zinc sulfate approximately 30-fold, added putrescine and linoleic acid, and increased several amino acids and vitamins (choline, myo-inositol) to achieve true protein-free clonal growth of CHO cells. See the Key Structural Differences section above for the full biochemical rationale.

Coon's Modification is specifically tailored for hybridoma production (virally fused hybrid cells via Sendai virus treatment) and culturing certain rodent cell lines. With appropriate supplements it is also used for rat thyroid cell lines such as FRTL-5.^[8] The 2X product pages list hybrid-cell culture, rat thyroid-cell culture, hormone-response and cell-signaling research, iodide-transport and thyroid-function research, cell-based chemical-response studies, chondrogenic differentiation research, and semisolid or agarose culture development. Ham's F-12 more broadly is used for CHO cells, cancer cells, primary cells, chicken embryonic cells, myeloma and hybridoma cells, primary rat hepatocytes, rat prostate epithelial cells, and normal rat kidney cells.

While lacking proteins, lipids, and growth factors, Ham's F-12 requires supplementation with serum or serum-free additives to facilitate optimal growth. Coon's modified F-12 contains no added proteins or growth factors; FRTL-5 thyroid cells, for example, are grown with serum and a defined hormone mixture.^[8]

The medium employs a sodium bicarbonate buffer system, necessitating a 5-10% CO₂ environment to maintain physiological pH. For the bicarbonate-containing 2X variants, use a CO₂-controlled incubator once diluted to 1X (2,676 mg/L sodium bicarbonate at 1X; classically equilibrated with 10% CO₂, commonly 5%) and keep time in room air short. For the bicarbonate-free variants it depends on the buffer you add: with sodium bicarbonate, use a CO₂-controlled incubator; with a non-volatile buffer such as HEPES, CO₂ is not needed for pH control. Leaving sodium bicarbonate out lets the solution or matrix you mix with the 2X carry the buffer, or lets you set your own bicarbonate level for your CO₂ tension; pH and sodium bicarbonate concentration are critical to cell growth, so monitor pH closely.

HEPES is not a standard catalog option for this family, and no 2X variant contains HEPES, which keeps it out of HEPES-sensitive assays. 25 mM HEPES is available as a customization on request via support@diagnocine.com.

Yes. Every variant is quadruple-stage filtered to a 0.04 micron final pore size, which lowers the risk of clogging microchannels, valves, and membranes in organ-on-a-chip, hydrogel-based 3D, and microfluidic perfusion systems.^[13,14,15]

Yes. Standard concentration for this family is 2X, with 4.0 g/L glucose (2.0 g/L at 1X) and 25 mM HEPES as a customization option. Other concentrations, additions of chemicals, compounds, proteins, supplements, a different pH, and modifications are available on request via support@diagnocine.com. A 10 nm (0.01 micron) ultra-filtered grade for automated bioreactors, liquid-handling robots, and closed perfusion loops is also made on request — email your catalog number and volume.

All 2X variants are supplied in a 500 mL size. Store at 2-8°C, away from bright light; do not freeze. A lot-specific Certificate of Analysis is available on request at support@diagnocine.com.

References

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
Coon, H.G. & Weiss, M.C. (1969). A quantitative comparison of formation of spontaneous and virus-produced viable hybrids. Proc. Natl. Acad. Sci. USA, 62(3): 852-859. DOI: 10.1073/pnas.62.3.852
Ambesi-Impiombato, F.S., Parks, L.A. & Coon, H.G. (1980). Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc. Natl. Acad. Sci. USA, 77(6): 3455-3459. DOI: 10.1073/pnas.77.6.3455
Borowicz, S., Van Scoyk, M., Avasarala, S., et al. (2014). The soft agar colony formation assay. Journal of Visualized Experiments, (92): e51998. DOI: 10.3791/51998
Barnes, D. & Sato, G. (1980). Methods for growth of cultured cells in serum-free medium. Analytical Biochemistry, 102(2): 255-270. DOI: 10.1016/0003-2697(80)90151-7
Morton, H.J. (1970). A survey of commercially available tissue culture media. In Vitro, 6(2): 89-108. DOI: 10.1007/BF02616111
Berthois, Y., Katzenellenbogen, J.A. & Katzenellenbogen, B.S. (1986). 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, 83(8): 2496-2500. DOI: 10.1073/pnas.83.8.2496
Bhatia, S.N. & Ingber, D.E. (2014). Microfluidic organs-on-chips. Nature Biotechnology, 32(8): 760-772. DOI: 10.1038/nbt.2989
Low, L.A., Mummery, C., Berridge, B.R., Austin, C.P. & Tagle, D.A. (2021). Organs-on-chips: into the next decade. Nature Reviews Drug Discovery, 20(5): 345-361. DOI: 10.1038/s41573-020-0079-3
Leung, C.M., de Haan, P., Ronaldson-Bouchard, K., et al. (2022). A guide to the organ-on-a-chip. Nature Reviews Methods Primers, 2: 33. DOI: 10.1038/s43586-022-00118-6
Drexler, H.G. & Uphoff, C.C. (2002). Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention. Cytotechnology, 39(2): 75-90. DOI: 10.1023/A:1022913015916
Halldorsson, S., Lucumi, E., Gómez-Sjöberg, R. & Fleming, R.M.T. (2015). Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices. Biosensors and Bioelectronics, 63: 218-231. DOI: 10.1016/j.bios.2014.07.029

References [1]-[6] are the Coon's Modification family bibliography; references [7]-[17] are cited on the 2X product pages.

FluxMPS™ Platform

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.

FluxMPS™ Ham's F-12 Nutrient Mixture, Coon's Modification [2X] Family

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