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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 supports the growth of cells in cell culture by serving as an auxiliary energy source, especially when the 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 2). The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. 

pyrrolidone-carboxylic-acid

Figure 1. L-glutamine spontaneously decomposes into ammonia and pyrrolidone carboxylic acid at a rate dependent on pH and temperature.

controlled-delivery-glutamine
Figure 2. GlutaMAX supplement allows for controlled delivery of L-glutamine from media to cells in culture.
stability-of-gmax
Figure 3. L-glutamine degrades faster than GlutaMAX Supplement in media at 37°C. DMEM was supplemented with GlutaMAX Supplement or L-glutamine, dispensed into vials, and stored at 37°C. Samples were taken daily and frozen at –20°C. Levels of GlutaMAX Supplement and L-glutamine were determined by HPLC.
ammonia-levels
Figure 4. Ammonia levels in supplemented media. DMEM was supplemented with GlutaMAX Supplement or L-glutamine, dispensed into vials, and stored at 37°C. Samples were taken daily and frozen at –20°C. Levels of ammonia were determined by HPLC.


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 5. 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 6. 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 7 and 8 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 9 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 7. 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 8. IgG1 production. The samples from Figure 7 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 9. 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.


Description		Classical medium with L-glutamineClassical medium without L-glutamineQuantityMedium with GlutaMAX Supplement
 Cat. No.Cat. No.QuantityCat. No.Quantity
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
Low glucose, contains sodium pyruvate
11885076 1,000 mL10567014
(in EU 21885025)		500 mL
11885084 500 mL
11885092 10 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains sodium pyruvate		11995040 1,000 mL10569010
(in EU 31966021)		500 mL
11995081 6 x 1,000 mL
1199506510313021500 mL
11995073 10 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains no sodium pyruvate		11965084119600511,000 mL10566016
(in EU 61965026)		500 mL
11965126119600776 x 1,000 mL
1196509211960044500 mL
119651181196006910 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains HEPES buffer but no sodium pyruvate		12430047 1,000 mL10564011
(in EU 32430027)		500 mL
12430054 500 mL
12430062 10 x 500 mL
DMEM/F-12 (1X), liquid, 1:111320033 500 mL10565018
(in EU 31331028)		500 mL
Ham’s F-12 Nutrient Mix (1X), liquid11765047 1,000 mL31765035
(in EU 31765027)		500 mL
11765070 6 x 1,000 mL
11765054 500 mL
11765062 10 x 500 mL
Iscove’s Modified Dulbecco’s Medium (IMDM) (1X), liquid12440046 1,000 mL31980030
(in EU 31980022)		500 mL
12440053 500 mL
12440061 10 x 500 mL
Minimum Essential Medium (MEM) alpha (1X), liquid
Contains no ribonucleosides or deoxyribonucleosides		12561049 1,000 mL32561037
(in EU 32561029)		500 mL
12561056 500 mL
Minimum Essential Medium (MEM) alpha (1X), liquid
Contains ribonucleosides and deoxyribonucleosides		12571048 1,000 mL32571036
(in EU 32571028)		500 mL
12571063 500 mL
12571071 10 x 500 mL
Minimum Essential Medium (MEM), liquid
Contains Earle’s salts		11095072110900731,000 mL41090036
(in EU 41090028)		500 mL
1109508011090081500 mL
110950981109009910 x 500 mL
Minimum Essential Medium (MEM), liquid
Contains Earle’s salts and HEPES buffer		 12360038500 mL42360032
(in EU 42360024)		500 mL
Opti-MEM I Reduced-Serum Medium (1X), liquid31985062 100 mL51985034
(in EU 51985026)		500 mL
31985070 500 mL
31985088 1,000 mL
RPMI 1640 Medium (1X), liquid11875085218700841,000 mL61870036
(in EU 61870010)		500 mL
11875135 6 x 1,000 mL
1187509321870076500 mL
118751192187009210 x 500 mL
11875101 100 mL
11875127 20 x 100 mL
RPMI 1640 Medium (1X), liquid
Contains HEPES buffer		22400071 1,000 mL72400047
(in EU 72400021)		500 mL
22400089 500 mL
22400105 10 x 500 mL
    GlutaMAX Supplement
 Cat. No. QuantityCat. No.Quantity
L-Glutamine 200 mM (100X), liquid25030149 20 mL35050061
(in EU 35050038)		100 mL
25030081 100 mL
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 2). The L-glutamine and L-alanine can then be taken up by the cells and utilized for protein production or in the TCA cycle. 

pyrrolidone-carboxylic-acid

Figure 1. L-glutamine spontaneously decomposes into ammonia and pyrrolidone carboxylic acid at a rate dependent on pH and temperature.

controlled-delivery-glutamine
Figure 2. GlutaMAX supplement allows for controlled delivery of L-glutamine from media to cells in culture.
stability-of-gmax
Figure 3. L-glutamine degrades faster than GlutaMAX Supplement in media at 37°C. DMEM was supplemented with GlutaMAX Supplement or L-glutamine, dispensed into vials, and stored at 37°C. Samples were taken daily and frozen at –20°C. Levels of GlutaMAX Supplement and L-glutamine were determined by HPLC.
ammonia-levels
Figure 4. Ammonia levels in supplemented media. DMEM was supplemented with GlutaMAX Supplement or L-glutamine, dispensed into vials, and stored at 37°C. Samples were taken daily and frozen at –20°C. Levels of ammonia were determined by HPLC.


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 5. 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 6. 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 7 and 8 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 9 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 7. 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 8. IgG1 production. The samples from Figure 7 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 9. 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.

Media cross-reference

Description		Classical medium with L-glutamineClassical medium without L-glutamineQuantityMedium with GlutaMAX Supplement
 Cat. No.Cat. No.QuantityCat. No.Quantity
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
Low glucose, contains sodium pyruvate
11885076 1,000 mL10567014
(in EU 21885025)		500 mL
11885084 500 mL
11885092 10 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains sodium pyruvate		11995040 1,000 mL10569010
(in EU 31966021)		500 mL
11995081 6 x 1,000 mL
1199506510313021500 mL
11995073 10 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains no sodium pyruvate		11965084119600511,000 mL10566016
(in EU 61965026)		500 mL
11965126119600776 x 1,000 mL
1196509211960044500 mL
119651181196006910 x 500 mL
Dulbecco’s Modified Eagle Medium (DMEM) (1X), liquid
High glucose, contains HEPES buffer but no sodium pyruvate		12430047 1,000 mL10564011
(in EU 32430027)		500 mL
12430054 500 mL
12430062 10 x 500 mL
DMEM/F-12 (1X), liquid, 1:111320033 500 mL10565018
(in EU 31331028)		500 mL
Ham’s F-12 Nutrient Mix (1X), liquid11765047 1,000 mL31765035
(in EU 31765027)		500 mL
11765070 6 x 1,000 mL
11765054 500 mL
11765062 10 x 500 mL
Iscove’s Modified Dulbecco’s Medium (IMDM) (1X), liquid12440046 1,000 mL31980030
(in EU 31980022)		500 mL
12440053 500 mL
12440061 10 x 500 mL
Minimum Essential Medium (MEM) alpha (1X), liquid
Contains no ribonucleosides or deoxyribonucleosides		12561049 1,000 mL32561037
(in EU 32561029)		500 mL
12561056 500 mL
Minimum Essential Medium (MEM) alpha (1X), liquid
Contains ribonucleosides and deoxyribonucleosides		12571048 1,000 mL32571036
(in EU 32571028)		500 mL
12571063 500 mL
12571071 10 x 500 mL
Minimum Essential Medium (MEM), liquid
Contains Earle’s salts		11095072110900731,000 mL41090036
(in EU 41090028)		500 mL
1109508011090081500 mL
110950981109009910 x 500 mL
Minimum Essential Medium (MEM), liquid
Contains Earle’s salts and HEPES buffer		 12360038500 mL42360032
(in EU 42360024)		500 mL
Opti-MEM I Reduced-Serum Medium (1X), liquid31985062 100 mL51985034
(in EU 51985026)		500 mL
31985070 500 mL
31985088 1,000 mL
RPMI 1640 Medium (1X), liquid11875085218700841,000 mL61870036
(in EU 61870010)		500 mL
11875135 6 x 1,000 mL
1187509321870076500 mL
118751192187009210 x 500 mL
11875101 100 mL
11875127 20 x 100 mL
RPMI 1640 Medium (1X), liquid
Contains HEPES buffer		22400071 1,000 mL72400047
(in EU 72400021)		500 mL
22400089 500 mL
22400105 10 x 500 mL
    GlutaMAX Supplement
 Cat. No. QuantityCat. No.Quantity
L-Glutamine 200 mM (100X), liquid25030149 20 mL35050061
(in EU 35050038)		100 mL
25030081 100 mL

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

Protect your cell cultures with Gibco Penicillin-Streptomycin-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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Order DMEM with GlutaMAX supplement
Order RPMI 1640 with GlutaMAX supplement
Order DMEM F-12 with GlutaMAX supplement
Order MEM with GlutaMAX supplement
Order other media with GlutaMAX supplement

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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