Glutamine

Manufactured by US Biological

Glutamine is a non-essential amino acid that plays a crucial role in various cellular processes. It serves as a building block for proteins and is involved in the synthesis of other amino acids, nucleic acids, and important biomolecules. Glutamine is a key nutrient for maintaining cellular function and supporting various physiological processes.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Pantothenate, by Merck Group (2 mentions) Pyruvate, by Merck Group (2 mentions) Glucose, by Thermo Fisher Scientific (2 mentions) Pantothenate, by US Biological (2 mentions) Epoch plate reader, by Agilent Technologies (2 mentions) Sulforhodamine b, by Merck Group (2 mentions) 24 well plate, by Greiner (2 mentions) Sodium pyruvate final, by Thermo Fisher Scientific (1 mentions) Red blood cell lysis buffer, by Merck Group (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) β mercaptoethanol final, by Merck Group (1 mentions) Tetracycline free fbs, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by US Biological (1 mentions) Dmem medium, by US Biological (1 mentions)

4 protocols using glutamine

Cell Culture Protocols for Various Cell Types

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Erythrocytes cells were depleted from
total mouse splenocytes using red blood cell lysis buffer (Sigma,
cat. R7757). Mouse lymphocytes were cultured in RPMI 1640 (cat. 11875;
Gibco) supplemented with 10% (v/v) inactivated FCS (Gibco), 0.0002%
β-mercaptoethanol final (Sigma, cat. M7522), penicillin 50 units
per liter–streptomycin 50 mg per liter (Sigma, cat. P4333),
1 mM sodium pyruvate final (Gibco, cat. 11360) nonessential amino
acids (Life Technologies, cat. 11140) and 2 mM glutamine (US Biological,
cat. G7120). HEK 293T cells were cultured in Dulbecco’s Modified
Eagle’s Medium supplemented with 10% (v/v) inactivated FBS
(Gibco). Saccharomyces cerevisiae strain W303 was
cultured in YPD medium. Toxoplasma gondii (RH strain)
tachyzoites were grown in human foreskin fibroblasts (HFF) cultured
in Dulbecco’s Modified Eagles Medium (DMEM; Invitrogen) supplemented
with 10% tetracycline-free FBS (HyClone), 2 mM glutamine, 10 mM HEPES
(pH 7.5), and 20 μg/mL gentamicin.

Swee L.K., Lourido S., Bell G.W., Ingram J.R, & Ploegh H.L. (2014). One-Step Enzymatic Modification of the Cell Surface Redirects Cellular Cytotoxicity and Parasite Tropism. ACS Chemical Biology, 10(2), 460-465.

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Metabolic Regulation of Treg Cells

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nT_reg cells were isolated as described above and cultured in either normal T cell medium, T cell medium lacking glucose (Thermo Fisher Scientific, catalog no. 11879020), or T cell medium lacking amino acids including glutamine (US Biological, catalog no. R9010-02) substituted with dialyzed FBS (Gibco, catalog no. A3382001) for 24 hours at 1 × 10⁵ cells per well. T cell medium included 2000 U of IL-2 and CD3/CD28 beads as described above. iT_reg cells were isolated and polarized as described above for 3 days. Following polarization, iT_reg cells were cultured in normal T cell medium or T cell medium lacking glucose or amino acids for 24 hours. Both nT_reg and iT_reg cells were then collected, and RNA was extracted as described above. For stability assays, cells were cultured as described above for 48 hours, then collected, and stained for Foxp3 for flow cytometry.

Montauti E., Weinberg S.E., Chu P., Chaudhuri S., Mani N.L., Iyer R., Zhou Y., Zhang Y., Liu C., Xin C., Gregory S., Wei J., Zhang Y., Chen W., Sun Z., Yan M, & Fang D. (2022). A deubiquitination module essential for Treg fitness in the tumor microenvironment. Science Advances, 8(47), eabo4116.

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HMEC-1 Cell Growth Assay

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HMEC-1 cells or HMEC-1 cells overexpressing xCT or SLC1A3 were seeded into 24-well plates (Greiner Bio-One) at a density of 6.5 x 10³ cells per well in 0.5 mL medium. The following day, medium was changed to 1 mL HMEC-1 growth medium with any indicated supplements. For assays with added aspartate, medium was changed to custom DMEM medium without glucose, glutamine, or pantothenate (US Biological) supplemented with 5.55 mM glucose (Gibco), 1 mM pyruvate (Sigma), and 25 μM pantothenate (Sigma) along with supplements above: EGF, hydrocortisone, glutamine, HI-FBS. Media was buffered with sodium hydroxide to pH 7.95. Designated plates were placed in hypoxia chamber at 1% oxygen. Cell number was measured at the indicated time points using sulforhodamine B (Sigma) staining, as previously described [42 (link)]. Absorbance at 510 nm was read on an Epoch plate reader (BioTek). Relative growth was determined as (treatment absorbance / control absorbance).

Cohen E.B., Geck R.C, & Toker A. (2020). Metabolic pathway alterations in microvascular endothelial cells in response to hypoxia. PLoS ONE, 15(7), e0232072.

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HMEC-1 Cell Growth Assay under Hypoxia

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HMEC-1 cells or HMEC-1 cells overexpressing xCT or SLC1A3 were seeded into 24-well plates (Greiner Bio-One) at a density of 6.5 x 10 3 cells per well in 0.5 mL medium. The following day, medium was changed to 1 mL HMEC-1 growth medium with any indicated supplements. For assays with added aspartate, medium was changed to custom DMEM medium without glucose, glutamine, or pantothenate (US Biological) supplemented with 5.55 mM glucose (Gibco), 1 mM pyruvate (Sigma), and 25 µM pantothenate (Sigma) along with supplements above: EGF, hydrocortisone, glutamine, HI-FBS. Media was buffered with sodium hydroxide to pH 7.95.
Designated plates were placed in hypoxia chamber at 1% oxygen. Cell number was measured at the indicated time points using sulforhodamine B (Sigma) staining, as previously described (42) .
Absorbance at 510 nm was read on an Epoch plate reader (BioTek). Relative growth was determined as (treatment absorbance / control absorbance).

Cohen E.B., Geck R.C., & Toker A. (2020). Metabolic pathway alterations in microvascular endothelial cells in response to hypoxia.

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