L-Glutamine & GlutaMAX Cell Culture Supplements

L-glutamine is an essential amino acid additive for cell culture

L-glutamine is an amino acid supplement commonly added to mammalian cell culture media. L-glutamine serves as an auxiliary energy source, especially when cells are rapidly dividing and also can be used by cells as a source of nitrogen for the synthesis of proteins, nucleic acids, etc.

In aqueous solutions such as cell culture media, L-glutamine decomposes into ammonia, which is toxic to cells. To minimize the effects of this, researchers can adopt a fed-batch strategy to continuously feed low levels of L-glutamine into the culture or they can use an alternative formulation of L-glutamine such as Gibco GlutaMAX supplement, designed to be more stable in solution.

L-glutamine for cell culture

L-glutamine is an essential nutrient in cell cultures for energy production as well as protein and nucleic acid synthesis. We offer this amino acid media supplement in both liquid and powdered forms. Powdered glutamine is supplied as a USP-grade chemical, and liquid formulations are supplied as a ready-to-use 200 mM stock solution.

GlutaMAX supplement keeps your cells healthier longer

Although L-glutamine is an essential nutrient in cell cultures, it spontaneously degrades in cell culture media generating ammonia and pyrrolidine carboxylic acid as byproducts. Gibco GlutaMAX supplement is an improved cell culture supplement that can be used as a direct substitute for L-glutamine in your cell culture media.

GlutaMAX supplement is a dipeptide, L-alanine-L-glutamine, which is more stable in aqueous solutions and does not spontaneously degrade.

Choose from a variety of formulations

We offer many formulations in which the GlutaMAX dipeptide substitutes for L-glutamine. Options include DMEM, MEM, RPMI, Opti-MEM and other basal media.

GlutaMAX supplement is also available as a stand-alone supplement. Use the 200 mM solution GlutaMAX supplement as a direct substitute for L-glutamine at equimolar concentrations in your current cell culture media formulation. GlutaMAX supplement is stable at room temperature meaning freeze-thaw cycles can be avoided, reducing the risk of culture contamination from frequent liquid handling steps.

Gibco GlutaMAX media and media supplement are suitable for both adherent and suspension mammalian cell cultures with little or no adaptation required.

Taking your work to the clinic? Be sure to check out CTS GlutaMAX Supplement

NOTE:  This supplement is suitable for mammalian cell cultures. It is not recommended for insect cell cultures.

GlutaMAX supplement vs. L-glutamine

Get the most from your cell cultures with GlutaMAX supplement

How does GlutaMAX supplement work?

The mechanism of dipeptide utilization involves the gradual release of peptidase during culture to allow the gradual hydrolysis of the dipeptide in the medium (Figure 1). This can be compared to the strategy of a fed-batch culture in which L-glutamine is continuously fed into the culture but maintained at low concentration. The result is an efficient energy metabolism and a high-growth yield.

Controlled delivery of L-glutamine

Figure 1. Controlled delivery of L-glutamine from media to cells in culture.

When cells are cultured in media that contains GlutaMAX supplement, the cells gradually release aminopeptidases which hydrolyze the dipeptide, slowly releasing L-alanine and L-glutamine into the culture media. The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. This can be compared to the fed-batch culture strategy in which L-glutamine is continuously fed into the culture but maintained at a low concentration. The result is an efficient energy metabolism and high growth yield without excess ammonia.

showcase.par.95467.image.192.128.1.row18-showcase-jpg

Figure 2. Human mesenchymal stem cells cultured with GlutaMAX media and stained with DAPI nuclear stain and primary CD105 antibody bound to a secondary Invitrogen Alexa Fluor 488 antibody.

l-glutamine.par.72420.image.200.128.1.

Figure 3.  Human mesenchymal stem cells cultured with GlutaMAX media under adipogenic differentiating conditions and stained with Hoechst 33342 nuclear stain, Invitrogen LipidTOX Green neutral lipid and Invitrogen MitoTracker Red.

How does GlutaMAX supplement improve your cell culture?

Using GlutaMAX supplement in place of L-glutamine in cell culture can improve cell viability and growth, potentially increasing productivity levels.

Results may vary depending upon the cell line used. In an application-specific example, Figures 4 and 5 show an AE-1 cell growth curve and recombinant IgG1 production. The GlutaMAX supplement culture demonstrates improved cell numbers and productivity.

GlutaMAX supplement can extend cell culture life, which may reduce the number of times the cells must be passaged and saves you time and money.  

Figure 6 compares MDBK cells cultured in D-MEM with 10% FBS and L-glutamine or GlutaMAX supplement. Cells cultured in GlutaMAX supplement reach peak density two days later and viability declines less rapidly than that observed in cultures with L-glutamine supplementation. The slight increase in lag phase is attributed to the time needed to release the peptidase and digest the dipeptide.  

This allows a gradual increase in availability of L-glutamine to the cells. (Appl Biochem Biotechnol 30:29 (1991)).

l-glutamine.par.97086.image.325.284.1.

Figure 4. Cell growth. AE-1 mouse myeloma cells were seeded at 1x105 cells/mL. Samples were taken daily after three days and evaluated in triplicate for cell density. Cell viability was determined by trypan blue exclusion. Cell density continued to increase in samples containing GlutaMAX supplement past the point when L-glutamine containing samples decreased in cell density.

l-glutamine.par.59356.image.325.283.1.

Figure 5. IgG1 production. The samples from Figure 4 were also examined by ELISA for IgG1 production. GlutaMAX supplement–containing samples outperformed L-glutamine–containing samples.

 

l-glutamine.par.68135.image.325.283.1.

Figure 6. MDBK cells were seeded at approximately 1x105 cells/flask in D-MEM with 10% FBS and L-glutamine or GlutaMAX supplement in 25 cm2 T-flasks.

How is L-glutamine used in cell culture?

L-glutamine is an essential amino acid required in cell culture media formulations. L-glutamine participates in the formation of purine and pyrimidine nucleotides, amino sugars, glutathione, L-glutamate, and other amino acids, as well as in protein synthesis and glucose production. Unlike most other amino acids, L-glutamine is not stable in solution. The rate at which degradation proceeds is a function of time, temperature, and pH. We offer Gibco GlutaMAX supplement as a more stable alternative to L-glutamine.

How should I thaw L-glutamine?

When L-glutamine is in a concentrated stock solution it easily precipitates when cooled. Warming the solution briefly in a 37°C water bath with gentle swirling will dissolve the precipitate. Do not use the product unless the precipitate is fully dissolved.

What is the shelf life of L-glutamine?

24 months from date of manufacture

How much L-glutamine should be added to media?

Gibco L-glutamine is supplied as a ready-to-use 200 mM stock solution that acts as a cell culture supplement. The optimal concentration is dependent upon the cell type and medium used to culture the cells, but generally falls in the range of 2–6 mM.

What is the difference between L-Glutamine and GlutaMAX supplement?

L-glutamine is an essential nutrient in cell cultures for energy production as well as protein and nucleic acid synthesis. However, L-glutamine in cell culture media spontaneously degrades, generating ammonia and pyrrolidine carboxylic acid as byproducts.

GlutaMAX supplement is a dipeptide, L-alanine-L-glutamine, which is more stable in aqueous solutions and does not spontaneously degrade. Gibco GlutaMAX media and media supplemented with GlutaMAX supplement are suitable for both adherent and suspension mammalian cell cultures with little or no adaptation required.

How does GlutaMAX supplement work?

The mechanism of dipeptide utilization involves the gradual release of peptidase during culture to allow the gradual hydrolysis of the dipeptide in the medium (Figure 1). This can be compared to the strategy of a fed-batch culture in which L-glutamine is continuously fed into the culture but maintained at low concentration. The result is an efficient energy metabolism and a high-growth yield.

Controlled delivery of L-glutamine

Figure 1. Controlled delivery of L-glutamine from media to cells in culture.

When cells are cultured in media that contains GlutaMAX supplement, the cells gradually release aminopeptidases which hydrolyze the dipeptide, slowly releasing L-alanine and L-glutamine into the culture media. The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. This can be compared to the fed-batch culture strategy in which L-glutamine is continuously fed into the culture but maintained at a low concentration. The result is an efficient energy metabolism and high growth yield without excess ammonia.

showcase.par.95467.image.192.128.1.row18-showcase-jpg

Figure 2. Human mesenchymal stem cells cultured with GlutaMAX media and stained with DAPI nuclear stain and primary CD105 antibody bound to a secondary Invitrogen Alexa Fluor 488 antibody.

l-glutamine.par.72420.image.200.128.1.

Figure 3.  Human mesenchymal stem cells cultured with GlutaMAX media under adipogenic differentiating conditions and stained with Hoechst 33342 nuclear stain, Invitrogen LipidTOX Green neutral lipid and Invitrogen MitoTracker Red.

How does GlutaMAX supplement improve your cell culture?

Using GlutaMAX supplement in place of L-glutamine in cell culture can improve cell viability and growth, potentially increasing productivity levels.

Results may vary depending upon the cell line used. In an application-specific example, Figures 4 and 5 show an AE-1 cell growth curve and recombinant IgG1 production. The GlutaMAX supplement culture demonstrates improved cell numbers and productivity.

GlutaMAX supplement can extend cell culture life, which may reduce the number of times the cells must be passaged and saves you time and money.  

Figure 6 compares MDBK cells cultured in D-MEM with 10% FBS and L-glutamine or GlutaMAX supplement. Cells cultured in GlutaMAX supplement reach peak density two days later and viability declines less rapidly than that observed in cultures with L-glutamine supplementation. The slight increase in lag phase is attributed to the time needed to release the peptidase and digest the dipeptide.  

This allows a gradual increase in availability of L-glutamine to the cells. (Appl Biochem Biotechnol 30:29 (1991)).

l-glutamine.par.97086.image.325.284.1.

Figure 4. Cell growth. AE-1 mouse myeloma cells were seeded at 1x105 cells/mL. Samples were taken daily after three days and evaluated in triplicate for cell density. Cell viability was determined by trypan blue exclusion. Cell density continued to increase in samples containing GlutaMAX supplement past the point when L-glutamine containing samples decreased in cell density.

l-glutamine.par.59356.image.325.283.1.

Figure 5. IgG1 production. The samples from Figure 4 were also examined by ELISA for IgG1 production. GlutaMAX supplement–containing samples outperformed L-glutamine–containing samples.

 

l-glutamine.par.68135.image.325.283.1.

Figure 6. MDBK cells were seeded at approximately 1x105 cells/flask in D-MEM with 10% FBS and L-glutamine or GlutaMAX supplement in 25 cm2 T-flasks.

How is L-glutamine used in cell culture?

L-glutamine is an essential amino acid required in cell culture media formulations. L-glutamine participates in the formation of purine and pyrimidine nucleotides, amino sugars, glutathione, L-glutamate, and other amino acids, as well as in protein synthesis and glucose production. Unlike most other amino acids, L-glutamine is not stable in solution. The rate at which degradation proceeds is a function of time, temperature, and pH. We offer Gibco GlutaMAX supplement as a more stable alternative to L-glutamine.

How should I thaw L-glutamine?

When L-glutamine is in a concentrated stock solution it easily precipitates when cooled. Warming the solution briefly in a 37°C water bath with gentle swirling will dissolve the precipitate. Do not use the product unless the precipitate is fully dissolved.

What is the shelf life of L-glutamine?

24 months from date of manufacture

How much L-glutamine should be added to media?

Gibco L-glutamine is supplied as a ready-to-use 200 mM stock solution that acts as a cell culture supplement. The optimal concentration is dependent upon the cell type and medium used to culture the cells, but generally falls in the range of 2–6 mM.

What is the difference between L-Glutamine and GlutaMAX supplement?

L-glutamine is an essential nutrient in cell cultures for energy production as well as protein and nucleic acid synthesis. However, L-glutamine in cell culture media spontaneously degrades, generating ammonia and pyrrolidine carboxylic acid as byproducts.

GlutaMAX supplement is a dipeptide, L-alanine-L-glutamine, which is more stable in aqueous solutions and does not spontaneously degrade. Gibco GlutaMAX media and media supplemented with GlutaMAX supplement are suitable for both adherent and suspension mammalian cell cultures with little or no adaptation required.

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