CHO CELLS MEDIA AND MEDIUM SUPPLEMENTS

The development of genetically-engineered cell lines to express therapeutically valuable recombinant proteins has created a major break-through in health care.

While the earliest methods of expressing recombinant proteins relied on bacterial systems, it is now recognized that the pharmacology and efficacy of recombinant proteins produced for therapeutic applications are affected by post-translational modifications such as glycosylation and phosphorylation. This has led to the use of higher order cell systems.

The Chinese Hamster Ovary cell line (CHO) was one of the first mammalian cell lines successfully developed for use in the production of therapeutically valuable proteins. Today, variations of this cell line are widely used for this purpose.  To accommodate the variety among different CHO clones, Sigma offers several different formulations to meet the needs of specific end users.

For more information on medium selections click here

• Liquid
• Kit consists of 6 proprietary medium formulations containing inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, recombinant human insulin, plant hydrolysates, other organic compounds, trace elements, and surfactants.
• Medium formulations do not contain antibiotics, antimycotics, L-glutamine, or transferrin.
• Animal Component-free
• Sterile filtered
• Cell culture tested

To determine which medium is optimal for an individual CHO cell clone, it is often necessary to screen multiple formulations. This kit provides a platform from which the user can screen several unique CHO modifications. Initial screening of the six media for growth and productivity may offer enough improvement over your current culture system to suffice. However, if further medium optimization is desired, the user can select the top three performing media based on cell growth or productivity. These three formulations can be combined in mixture experiments as described in the kit and can be analyzed to determine a best-fit media based on growth and productivity.

• Liquid
• Proprietary formulation containing inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, recombinant human insulin, plant hydrolysates, other organic compounds, trace elements, and surfactants.
• Does not contain antibiotics, antimycotics, L-glutamine, or transferrin.
• Animal Component-free
• Sterile filtered
• Cell culture tested

This animal component-free medium (containing no hypoxanthine or thymidine) is formulated to maximize cell growth and recombinant protein production using the Dihydrofolate Reductase (DHFR) gene amplification system in DHFR- Chinese Hamster Ovary (CHO) cells.

This medium supports rapid initial cell growth and high levels of protein expression in suspension cultures. This medium supports and maintains high cell densities for extended periods, resulting in increased productivity.

• Liquid
• Proprietary formulation
• With inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, rHu-Insulin, other organic compounds, trace elements, and surfactants.
• With plant hydrolysates
• Without antibiotics, anti-mycotics,  transferrin, or L-glutamine: add 20-40 mL of 200 mM L-glutamine per liter,  prior to use.
• Without hypoxanthine and thymidine;
• pH 7.1-7.7
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Optimized for the production of recombinant proteins by cultured CHO cells, under protein-free and animal derived component-free conditions; designed to support the rapid growth, high cell densities and extended viability of genetically engineered CHO cells in suspension culture.

This formulation eliminates regulatory concerns about the risks of biological contamination of recombinant proteins that are associated with the use of animal components during cell culture.

May be used in Dihydrofolate Reductase (DHFR) gene amplification systems.

• Powder
• Proprietary formulation
• Formula contains inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, rHu-Insulin, other organic compounds, trace elements, and surfactants.
• Formula contains plant hydrolysates
• Formula without antibiotics, anti-mycotics, hypoxanthine, thymidine or transferrin.
• Without L-glutamine and sodium bicarbonat: supplement with 0.584 to 1.168 gm/L of L-glutamine and 1.5 gm/L of sodium bicarbonate.
• pH 6.0 +/- 1.0 without sodium bicarbonate, and 7.1-7.7 fully formulated.
• Formulated at 21.5 grams per liter of medium.
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

This is the powder version of C 5467.  It is only available as a custom formulation.

Optimized for the production of recombinant proteins by cultured CHO cells, under protein-free and animal derived component-free conditions; designed to support the rapid growth, high cell densities and extended viability of genetically engineered CHO cells in suspension culture.

This formulation eliminates  regulatory concerns about the risks of biological contamination of recombinant proteins that are associated with the use of animal components during cell culture.

May be used in Dihydrofolate Reductase (DHFR) gene amplification systems.

• Liquid
• Proprietary formulation
• With inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, rHu-Insulin, other organic compounds, trace elements, and surfactants.
• With plant hydrolysates
• Without antibiotics, phenol red, and L-glutamine: add 20 mL of 200 mM L-glutamine per liter,  prior to use.
• Without hypoxanthine and thymidine;
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Modified version of C 5467,  divalent cation concentrations have been optimized to promote better attachment. Designed to support the growth of, and expression of recombinant proteins by, adherent CHO cells in a variety of systems, such as; T-flasks, roller bottles and bioreactors using fiber discs or micro-carrier beads.

This formulation eliminates regulatory concerns about the risks of biological contamination of recombinant proteins that are associated with the use of animal components during cell culture.

May be used in Dihydrofolate Reductase (DHFR) gene amplification systems.

• Liquid
• Proprietary formulation
• With inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, rHu-insulin, other organic compounds, trace elements, and surfactants.
• Without plant hydrolysates
• Without antibiotics, anti-mycotics, transferrin, any undefined nutrients or supplement and L-glutamine: add 20-40 mL of 200 mM L-glutamine per liter, prior to use.
• Without hypoxanthine and thymidine;
• pH 7.1-7.7
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

All components in this animal protein free medium  are chemically defined. This allows biotechnologists to have greater control over the biological risk factors associated with the production of CHO expressed proteins; AND to optimize cell growth, protein production, and  the down-stream purification process.  Furthermore, this formulation offers the biotechnologist the best opportunity to manage and control lot-to-lot variations in productivity.

This formulation eliminates regulatory concerns about the risks of biological contamination of recombinant proteins that are associated with the use of animal components during cell culture.

May be used in Dihydrofolate Reductase (DHFR) gene amplification systems.

• Liquid
• With fetuin, insulin, BSA, transferrin, and HEPES and sodium bicarbonate buffers;
• Without L-glutamine: add 10.3 mL of 200 mM L-glutamine per liter, prior to use
• Without hypoxanthine and thymidine;
• Protein content: 100 ug/mL
• Sterile filtered
• Endotoxin tested
• Cell culture tested

Optimized for the production of recombinant proteins, under serum-free conditions, which are expressed in either suspension or attached cultured CHO cells.

This formulation reduces regulatory concerns, about the risks of biological contamination of recombinant therapeutics, that are associated with the use of serum.

May be used in Dihydrofolate Reductase (DHFR) gene amplification systems.

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Chinese Hamster Ovary (CHO) cell expression systems are complex, and are typically modified (or optimized) to meet the specific priorities and needs of the biotechnologist.  In addition to offering the above varieties of CHO media, Sigma provides supplements that you may find useful in your culture system. The table below identifies some of the most useful supplements for your evaluation and use.

Other important potential supplements such as the transferrins, albumins, fetuins, and protein hydrolysates are offered by Sigma in many varieties.  Please use the Sigma search engine to find these product families.

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Proprietary medium supplement specially formulated to boost cell growth for enhanced protein expression. May be used in combination with Polyamine Supplement (P 8483)

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Animal-component free. Proprietary aqueous mixture of fatty acids that have been found to be optimal for cell growth and protein expression.

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Prepared in cell culture grade water. Useful for supplementing liquid CHO media.

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

The glutamine synthetase (GS) system is used to express recombinant proteins from mammalian cells.

Glutamine-dependent cells transfected with a glutamine synthetase gene can be selectively grown in glutamine-deficient medium.

• Liquid, in DMSO
• Contains 4-(2-aminoethyl) benzene sulfonyl fluoride (AEBSF), pepstatin A, trans epoxysuccinyl L-leucylamido(4-guanidino) butane (E-64), bestatin, leupeptin, and aprotinin.
• Contains no metal chelators.

Endogenous enzymes, notably proteases and phosphatases, can degrade proteins. The best way to improve the yield of fully functional proteins is to add inhibitors of these lytic enzymes. This formulation contains inhibitors of serine, cysteine, and aspartic proteases, as well as inhibitors of aminopeptidases.

This cocktail may be used effectively to increase the quality of protein expressed in CHO cells. However, it should always be thoroughly tested for specific cell culture applications.

• Liquid
• Cell culture tested.

A media additive often used to protect cells growing in suspended culture systems from damage which may occur during agitation and aeration.

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Medium supplement formulated to boost cell growth for enhancement of protein expression.

• Powder
• Molecular Weight 84.01
• Purity >99.5%
• Cell culture tested.

Use to supplement cell culture media.

• Liquid
• Sterile filtered.
• Endotoxin tested.
• Cell culture tested.

Use to supplement cell culture media.