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Cell Culture

GlutaMAX™ media and supplement

 We build healthier cells 

  • Improves cell viability and growth
  • Minimizes ammonia build-up
  • Keeps cells healthier longer

Left panel:  Human mesenchymal stem cells cultured with GlutaMAX™ media and stained with DAPI nuclear stain and primary CD105 antibody bound to a secondary Alexa Flour® 488 antibody.

GlutaMAX™ facts

GIBCO® GlutaMAX™ media is our standard trusted cell culture media but it contains a stabilized form dipeptide from L-glutamine, L-alanyl-L-glutamine, that prevents degradation and ammonia build-up even during long-term cultures.  

Extremely stable in aqueous solution, the L-alanyl-L-glutamine dipeptide will not degrade into ammonia in storage or incubation like L-glutamine. Read about the Stability of GlutaMAX™-I vs. L-Glutamine.

See below how GlutaMAX™ media and supplement can improve your cell culture, save you time and money.


  • 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.  This generates ammonia as a byproduct, which is toxic to the cells2 and can affect protein glycosylation3,4 and cell viability, lowering protein production and changing glycosylation patterns.

    L-glutamine Graph Figure 1.  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™-I past the point when L-glutamine containing samples decreased in cell density.

    L-glutamine Graph Figure 2.  IgG1 Production.  The samples from figure 1 were also examined by ELISA for IgG1 production.  GlutaMAX™-I containing samples outperformed L-glutamine containing samples.


    Using GlutaMAX™-I 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 1 and 2 show an AE-1 cell growth curve and recombinant IgG1 production.  The GlutaMAX™-I culture demonstrates improved cell numbers and productivity.


  • Lower ammonia concentrations can be advantageous in attaining high cell yields, particularly for cells that are sensitive to ammonia toxicity5.      Cells can be sensitive to ammonia even at non-toxic levels, creating artifacts.

    The GlutaMAX™ dipeptide is split by aminopeptidases, releasing L-glutamine and L-alanine from the dipeptide.


    L-glutamine Chart Figure 3. Controlled delivery of L-glutamine from media to cells in culture.  


    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 3).  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.


  • L-glutamine Figure 4.  MDBK cells were seeded at approximately 1x105 cells/flask in D-MEM with 10% FBS and L-glutamine or GlutaMAX™-I in 25 cm2 T-flasks.

    GlutaMAX™-I 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 4     compares MDBK cells cultured in D-MEM with 10% FBS and L-glutamine or GlutaMAX™-I supplement.  Cells cultured in GlutaMAX™-I 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.2

Suitable for numerous applications

GlutaMAX™ media and media supplemented with GlutaMAX™-I are suitable for both adherent and suspension mammalian cell cultures including:



  • Culture systems requiring long periods of incubations without feeding (e.g. cloning assays)

  • Long-term studies requiring optimum standardization of media (e.g. cancer cell lines, long-term cultures passaged over time, toxicity testing)

  • Culture systems sensitive to ammonia (e.g. high-density bioreactors)
 
Human Stem Cells (HMS)

Figure 5:  Human mesenchymal stem cells cultured with GlutaMAX™ media under adipogenic differentiating conditions and stained with Hoechst 33342 nuclear stain, LipidTOX™ Green neutral lipid and MitoTracker® Red.


Common cell lines cultured with GlutaMAX-I™
MDBK Bovine KidkeyMDCK Canine Kidney 
HELA Human OvaryPer. C6 Human Embryonic Retinoblastoma 
298 Human Embryonic KidneyBHK Hamster Kidney 
AE-1 Mouse Hybridoma 3D9 Mouse Hybridoma 
CHO Hamster Ovary For a complete list of citations see references below 

Choose from a variety of formulations

We offer many widely used formulations in which the GlutaMAX™ dipeptide substitutes for L-glutamine.  They include D-MEM, MEM, RPMI, Opti-MEM and others.  You can also purchase the GlutaMAX™ dipeptide as a stand-alone supplement.  Use the 200 mM solution GlutaMAX™-I supplement as a direct substitute for L-glutamine at equimolar concentrations in your current cell culture media formulation.

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

References

  1. Tritsch, G.L. and Moore, G.E. (1962) Spontaneous Decomposition of Glutamine in Cell Culture Media.  Experimental Research 28, 360–364.
  2. Hassell, T., Gleave, S., and Butler, M. (1991) Growth Inhibition in Cell Culture.  Applied Biochemistry and Biotechnology 30, 30–41.
  3. Yang, M. and Butler, M. (2002) Effects of Ammonia and Glucosamine on the Heterogeneity of Erythropoietin Glycoforms.  Biotechnology Progress 18, 129–138.
  4. Yang, M. and Butler, M. (2000) Effects of Ammonia on the Glycosylation of Human Recombinant Erythropoietin in Culture.  Biotechnology Progress 16, 751–759.
  5. Christie, A. and Butler, M. (1994) Growth and Metabolism of a Murine Hybridoma in Cultures Containing Glutamine-based Dipeptides. FOCUS® 16, 1, 9.
  6. Brand, K., Feki, W., Hintzentern, J., von Langer, K., Luppa, P., and Schroener, C. (1989) Metabolism 38, 29.
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