Glutamine

Manufactured by Eurobio Scientific

Sourced in France

Glutamine is a fundamental amino acid that serves as a key substrate in cellular metabolism. It plays a crucial role in various physiological processes, including nitrogen transport, energy production, and cell growth and proliferation.

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L-glutamine (42 citations)

5 protocols using glutamine

Molecular Mechanisms of HMGB1 Signaling

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Cell culture media, cell transfection opti-MEM (1×) medium, epithelial growth factor (PHG0311), fetal bovine serum (FBS), insulin transferrin selenium, Pierce BCA Protein Assay Kit (23225), Lipofectamine 3000 Transfection Reagent (L3000008), and Lipofectamine RNAiMAX Transfection Reagent (13778150) were obtained from ThermoFisher Scientific (Illkirch-Graffenstaden, France). Antibiotics (streptomycin, penicillin, and amphotericin B) and glutamine were purchased from Eurobio Scientific (Les Ulis, France). Dimethyl sulfoxide (DMSO), HMGB1 (SRP6265, a mixture of different forms), and Fibronectin (F1141) were purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Human S100A4 (4137-S4), S100A8 (9876-S8), S100A9 (9254-S9), and MCP1 (279-MC-010/CF) recombinant proteins were obtained from R&D systems (Noyal-Châtillon-sur-Seiche, France). RAP (RAGE inhibitor, 553031) and TAK-242 (TLR4 inhibitor, 243984-11-4) were purchased from Merck (Saint-Quentin-Fallavier, France). LightCycler 480 SYBR Green I Master (04887352001) was provided by Roche (Meylan, France).

Henrioux F., Navel V., Belville C., Charnay C., Antoine A., Chiambaretta F., Sapin V, & Blanchon L. (2023). Inflammation of Dry Eye Syndrome: A Cellular Study of the Epithelial and Macrophagic Involvement of NFAT5 and RAGE. International Journal of Molecular Sciences, 24(13), 11052.

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Cellular Oxidative Stress Assays

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Fetal bovine serum (FBS) and glutamine were purchased from Eurobio^® (Les Ulis, France). Lipopolysaccharide from Escherichia coli K12 (LPS-EK) was obtained from InvivoGen^® (San Diego, California, United States) and inorganic phosphate (Pi) from Merck^® (Darmstadt, Germany). Exosome-free FBS, TRIzol^™ Reagent, RNase/DNase-free water, High-Capacity RNA-to-cDNA^™ kits, BCA^™ Protein Assay kits, and dihydroethidium (DHE) were purchased from Thermo Fisher Scientific^® (Waltham, Massachusetts, United States). Takyon^™ was obtained from Eurogentec^® (Liège, Belgium). 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) was purchased from Molecular Probes^® (Eugene, Oregon, United States). All other molecular and biochemical reagents were obtained from Sigma-Aldrich^® (Saint-Louis, Missouri, United States).

Yaker L., Tebani A., Lesueur C., Dias C., Jung V., Bekri S., Guerrera I.C., Kamel S., Ausseil J, & Boullier A. (2022). Extracellular Vesicles From LPS-Treated Macrophages Aggravate Smooth Muscle Cell Calcification by Propagating Inflammation and Oxidative Stress. Frontiers in Cell and Developmental Biology, 10, 823450.

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Toxin Effects on Caco-2 Oxygen Consumption

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The acute effect of toxins on oxygen consumption of Caco-2 cells was assessed using an XF24 extracellular flux analyzer (Seahorse Bioscience, North Billerica, USA). The procedure was performed according to the manufacturer’s instructions. Briefly, cells were cultured in XF24 cell culture microplates at 1.5 × 10⁴ cells per well and maintained as described above. Oxygen consumption rates (OCR) were measured in proliferated Caco-2 cells in non-buffered DMEM (Seahorse Bioscience) supplemented with 10 mM glucose, 2 mM sodium pyruvate (Sigma-Aldrich) and 2 mM glutamine (Eurobio) and adjusted to pH 7.4. OCR was monitored every 20 minutes before (basal level) and after injection of diluent or toxins (10 μM). OCR values from each well were normalized against viable cell counts (calculated with a Malassez cell) and expressed as a percentage of the baseline value.

Pierron A., Mimoun S., Murate L.S., Loiseau N., Lippi Y., Bracarense A.P., Schatzmayr G., He J.W., Zhou T., Moll W.D, & Oswald I.P. (2016). Microbial biotransformation of DON: molecular basis for reduced toxicity. Scientific Reports, 6, 29105.

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Isolation and Culture of CD34+ Progenitor Cells

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CD34⁺ progenitor cells were isolated by positive selection kit (MiltenyiBiotec) using immunomagnetic beads cell‐sorting system (AutoMACS; MiltenyiBiotec) from healthy leukapheresis samples (from allogenic stem cell transplantation donors), in agreement with our institutional ethics committee. CD34⁺ cells were cultivated as recently described²³ from day (D)1 to D6 in Iscove's modified Dulbecco's medium (IMDM) (Biochrom) supplemented with glutamine (Eurobio), containing 2% human AB serum (H2B), 100 IU/ml penicillin (Eurobio), 100 μg/ml streptomycin (Eurobio), 500 μg/ml human holo‐transferrin (Sigma‐Aldrich), 10 μg/ml recombinant human insulin (Sigma‐Aldrich), 2 IU/ml heparin (Braun), 100 ng/ml human stem cell factor (SCF) (MiltenyiBiotec) and 3 IU/ml erythropoietin (EPO) (Roche). From D7 to D20, cells were culture in the same medium but after SCF removal. UT7/EPO cells were maintained in α‐minimum essential medium (MEM) (Dominique Dutscher) supplemented with 10% FCS (Eurobio) and 2 IU/ml EPO as recently described.²⁴

Vong P., Messaoudi K., Jankovsky N., Gomilla C., Demont Y., Caulier A., Jedraszak G., Demagny J., Djordjevic S., Boyer T., Marolleau J.P., Rochette J., Ouled‐Haddou H, & Garçon L. (2022). HDAC6 regulates human erythroid differentiation through modulation of JAK2 signalling. Journal of Cellular and Molecular Medicine, 27(2), 174-188.

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Transient Transfection of HEK293FT Cells

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HEK293FT cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% (v/v) foetal bovine serum, 100 U/ml penicillin, 0.1 mg/ml streptomycin, and 1 mM glutamine (Eurobio) at 37°C in 5% CO2. They were then transiently transfected using Metafectene PRO (Biontex) following the manufacturer’s instructions.

Laboute T., Gandía J., Pellissier L.P., Corde Y., Rebeillard F., Gallo M., Gauthier C., Léauté A., Diaz J., Poupon A., Kieffer B.L., Le Merrer J, & Becker J.A. (2020). The orphan receptor GPR88 blunts the signaling of opioid receptors and multiple striatal GPCRs. eLife, 9, e50519.

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