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Choose Gibco L-glutamine for your cell culture

L-glutamine is an important amino acid supplement commonly added to mammalian cell culture media. L-glutamine serves as an auxiliary energy source, especially when cells are rapidly dividing. L-glutamine is also important in the production of purine and pyrimidine nucleotides, amino sugars, glutathione, L-glutamate, other amino acids, and plays a role in protein synthesis and glucose production.

Gibco L-glutamine is offered in both liquid and powdered forms. Powdered L-glutamine is supplied as a USP-grade chemical, and liquid formulations are supplied as ready-to-use 200 mM stock solutions. The optimal concentration of L-glutamine for cell culture is dependent upon the cell type and medium used, but generally falls in the range of 2–6 mM.

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Enable improved cell viability with GlutaMAX supplement

Explore the differences between L-glutamine and GlutaMAX supplement with the function and performance data provided below. Learn how you can get the most from your cell cultures with GlutaMAX supplement.

L-glutamine vs GlutaMAX supplement

In aqueous solutions such as cell culture media, L-glutamine spontaneously degrades, generating toxic ammonia and pyrrolidine carboxylic acid as byproducts. The rate at which degradation of L-glutamine proceeds is a function of time, temperature, and pH. This L-glutamine degradation can be exacerbated as pH levels rise in the presence of bases like phosphate or bicarbonate. Likewise, L-glutamine degradation can occur at acidic pH levels. To minimize the toxic effects of L-glutamine degradation on your cells, you can adopt a fed-batch strategy to continuously feed low levels of L-glutamine into the culture.

In contrast to standard L-glutamine, which is more sensitive to environmental factors such as pH, Gibco GlutaMAX supplement is a dipeptide, L-alanyl-L-glutamine, which is more stable in aqueous solutions and does not spontaneously degrade. Instead, cells gradually release aminopeptidases that hydrolyze the dipeptide, slowly releasing L-alanine and L-glutamine into the culture media (Figure 1). The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. 

Diagram showing that L-glutamine degrades into PCA and ammonia in cell culture, whereas GlutaMAX supplement allows for more controlled delivery of L-glutamine into cells without generation of these toxic byproducts

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

Using GlutaMAX supplement in cell culture

Gibco GlutaMAX supplement can be used as a direct substitute for L-glutamine at equimolar concentrations in your cell culture media, resulting in efficient energy metabolism and high-growth yield, without the detrimental effects of excess ammonia on your cells.

Microscopic view of fluorescently stained MSCs indicating CD105

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

Microscopic view of fluorescently stained MSCs in adipogenic differentiating conditions

Figure 3. Human mesenchymal stem cells cultured with GlutaMAX media under adipogenic differentiating conditions. They are 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 cell culture in place of L-glutamine can improve cell viability and growth, potentially increasing productivity levels. Figures 4 and 5 show an AE-1 cell growth curve and recombinant IgG1production. The GlutaMAX supplement culture demonstrates improved cell viability and productivity.

In addition, GlutaMAX supplement can extend the life of your cell culture, thus reducing the number of times the cells must be passaged, saving you time and money. Figure 6 compares MDBK cells cultured in DMEM 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, allowing a gradual increase in availability of L-glutamine to the cells. (Appl Biochem Biotechnol 1991, 30:29.)

Bar graph of viable cells vs time, showing GlutaMAX supplement supported increased cell density compared to other media formulations

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.

Bar graph of IgG1 vs time, showing GlutaMAX supplement supported increased IgG1 production compared to other media formulations

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.

 

Line graph of cell count vs time, showing GlutaMAX supplement supported better growth over time than L-glutamine

Figure 6. Cell count. 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.

Function

L-glutamine vs GlutaMAX supplement

In aqueous solutions such as cell culture media, L-glutamine spontaneously degrades, generating toxic ammonia and pyrrolidine carboxylic acid as byproducts. The rate at which degradation of L-glutamine proceeds is a function of time, temperature, and pH. This L-glutamine degradation can be exacerbated as pH levels rise in the presence of bases like phosphate or bicarbonate. Likewise, L-glutamine degradation can occur at acidic pH levels. To minimize the toxic effects of L-glutamine degradation on your cells, you can adopt a fed-batch strategy to continuously feed low levels of L-glutamine into the culture.

In contrast to standard L-glutamine, which is more sensitive to environmental factors such as pH, Gibco GlutaMAX supplement is a dipeptide, L-alanyl-L-glutamine, which is more stable in aqueous solutions and does not spontaneously degrade. Instead, cells gradually release aminopeptidases that hydrolyze the dipeptide, slowly releasing L-alanine and L-glutamine into the culture media (Figure 1). The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. 

Diagram showing that L-glutamine degrades into PCA and ammonia in cell culture, whereas GlutaMAX supplement allows for more controlled delivery of L-glutamine into cells without generation of these toxic byproducts

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

Using GlutaMAX supplement in cell culture

Gibco GlutaMAX supplement can be used as a direct substitute for L-glutamine at equimolar concentrations in your cell culture media, resulting in efficient energy metabolism and high-growth yield, without the detrimental effects of excess ammonia on your cells.

Microscopic view of fluorescently stained MSCs indicating CD105

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

Microscopic view of fluorescently stained MSCs in adipogenic differentiating conditions

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

Performance

How does GlutaMAX supplement improve your cell culture?

Using GlutaMAX supplement in cell culture in place of L-glutamine can improve cell viability and growth, potentially increasing productivity levels. Figures 4 and 5 show an AE-1 cell growth curve and recombinant IgG1production. The GlutaMAX supplement culture demonstrates improved cell viability and productivity.

In addition, GlutaMAX supplement can extend the life of your cell culture, thus reducing the number of times the cells must be passaged, saving you time and money. Figure 6 compares MDBK cells cultured in DMEM 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, allowing a gradual increase in availability of L-glutamine to the cells. (Appl Biochem Biotechnol 1991, 30:29.)

Bar graph of viable cells vs time, showing GlutaMAX supplement supported increased cell density compared to other media formulations

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.

Bar graph of IgG1 vs time, showing GlutaMAX supplement supported increased IgG1 production compared to other media formulations

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.

 

Line graph of cell count vs time, showing GlutaMAX supplement supported better growth over time than L-glutamine

Figure 6. Cell count. 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.

GlutaMAX supplement composition

We offer the GlutaMAX supplement as a standalone 200 mM solution of the L-alanyl-L-glutamine dipeptide, which can be used 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 you can avoid freeze-thaw cycles and decrease contamination risk from frequent liquid-handling steps.

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

GlutaMAX media formulations

In addition to the standalone supplement, you may select from many media formulations in which the GlutaMAX dipeptide is substituted for L-glutamine. These options include DMEM, MEM, RPMI, Opti-MEM and other basal media. The Gibco GlutaMAX supplement and media are suitable for both adherent and suspension mammalian cell cultures, with little to no further adaptation required.

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

Frequently asked questions (FAQs) for L-glutamine and GlutaMAX supplement

Would you like to learn more about using L-glutamine and GlutaMAX supplement in your cell culture studies?
Visit our L-glutamine and GlutaMAX FAQs

Product literature for L-glutamine and GlutaMAX supplement

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Gibco cell culture products are manufactured in facilities compliant with current good manufacturing practices (GMP) and adhere to a robust quality management system, helping to ensure the consistency, reliability, and high quality you can rely on.

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The 1 L Gibco bottle uses 38% less source material (32% less material for the 500 mL bottle) than comparable media bottles on the market today. This helps reduce the consumption of non-renewable resources, helping shrink your footprint.

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