L glutamine

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L-glutamine is an amino acid that is commonly used as a dietary supplement and in cell culture media. It serves as a source of nitrogen and supports cellular growth and metabolism.

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Glutamine (2902 citations) DMEM High Glucose with L-Glutamine (4 citations) L-glutamine (P/S/G; (4 citations) L-glutamine and pyruvate (3 citations) Penicillin/streptomycin/L-glutamine/heparin (3 citations) RPMI 1640 w/o L-glutamine (2 citations) FBS l-glutamine (2 citations) L-alanine-L-glutamine (2 citations) Ham’s F-12 w/L-glutamine (2 citations) GlutaMAX L-glutamine supplement (2 citations)

14 671 protocols using l glutamine

Establishing Docetaxel-Resistant Prostate Cancer Cell Lines

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The human prostate cancer cell lines PC3; C4-2B and LNCaP cells were grown in RPMI-1640 media supplemented with 10% fetal bovine serum (FBS), L-glutamine, nonessential amino acid, HEPES, 2mM L-glutamine and penicillin/streptomycin antibiotic solution (Fisher Scientific, Pittsburgh, PA). RWPE-1 cells were grown in keratinocyte serum-free medium (K-SFM), supplemented with 0.05 mg/ml bovine pituitary extracts (BPE), 5 ng/ml human recombinant epidermal growth factor (EGF), and penicillin/streptomycin antibiotic solution (Fisher Scientific, Pittsburgh, PA). All the cell lines were maintained in standard condition incubator at 37°C and 5% CO2.
Docetaxel resistant cells were prepared from the parental PC3 cells. The resistant cells were generated by gradually increasing the docetaxel concentration starting from 4nM to 40nM and were maintained in standard condition incubator for 8 months at 37°C and 5% CO2. The docetaxel resistant cells were labeled as PC3-R and were maintained in RPMI-1640 media supplemented with 10% fetal bovine serum (FBS), L-glutamine, nonessential amino acid, HEPES, 2mM L-glutamine and penicillin/streptomycin antibiotic solution (Fisher Scientific, Pittsburgh, PA).

Singh S.K., Lillard JW J.r, & Singh R. (2018). Reversal of drug resistance by planetary ball milled (PBM) nanoparticle loaded with resveratrol and docetaxel in prostate cancer. Cancer letters, 427, 49-62.

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Maintenance and Differentiation of Human Embryonic Stem Cells

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The hESC lines HSF-1 (UC01, 46XY), H1 (WA01, 46XY), H9 (WA09, 46XX) and UCLA1 (46XX) were maintained under self –renewal conditions on mouse embryonic fibroblast (MEF) layer in DMEM:F12 (Gibco BRL), 20% KnockOut Serum (Gibco BRL), 1% nonessential amino acids (NEAA, Gibco BRL), 1 mM L-glutamine (Gibco BRL), 0.1 mM β-mercaptoethanol (Gibco BRL), and 10ng/ml of basic fibroblast growth factor (FGF) from R&D. Undifferentiated hESC colonies were maintained as previously described [10 (link)]. Differentiation was performed on plates coated with growth factor reduced matrigel (BD Pharmigen) in DMEM:F12 supplemented with 20% FBS (Gibco BRL), 0.1 mM nonessential amino acids, 0.1 mM β-mercaptoethanol, 1 mM L-glutamine. Media was changed every 2 days during differentiation. For all experiments, hESCs were used between passages 35 and 50. All hESC experiments were conducted with prior approval from the UCLA Embryonic Stem Cell Research Oversight Committee. BJ fibroblast somatic cells were cultured in minimum essential medium (MEM) with Earle’s salt (Gibco BRL) and 1 mM L-glutamine, 10% FBS (Gibco BRL), 1% NEAA and 1 mM sodium pyruvate (Gibco BRL). Cells were passaged using 0.25% trypsin (Gibco BRL) every 7 days. HEK 293 FT cells were grown in DMEM High Glucose (Gibco BRL) supplemented with 10% FBS, 1× Pen-Strep (Gibco BRL), 1 mM L-glutamine and 1 mM sodium pyruvate.

Gkountela S., Li Z., Chin C.J., Lee S.A, & Clark A.T. (2014). PRMT5 is required for human embryonic stem cell proliferation but not pluripotency. Stem cell reviews, 10(2), 230-239.

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Culturing Neural Cells from Mouse Embryos

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Human embryonic kidney HEK293 cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Karlsruhe, Germany) supplemented with 10% fetal bovine serum (Gibco) and 10 mM L-glutamine (Gibco). The multipotent neural progenitor cell line C17.2 was maintained in DMEM supplemented with 10% fetal bovine serum, 5% horse serum (Gibco), and 10 mM L-glutamine. NSCs and neurons were separately established from cortex of embryonic day (E) 14-E16 C57BL/6 mice. Briefly, cortex was microdissected and stripped of meninges, and then tissues were mechanically dissociated into single-cell suspensions. For NSCs, cells were grown in DMEM/F-12 (Gibco) supplemented with 1 mM L-glutamine, 1% N2 supplement (Gibco), 20 μL/mL B-27 supplement minus vitamin A (Gibco), 100 μg/mL penicillin/streptomycin (Gibco), 20 ng/mL epidermal growth factor (EGF), and 20 ng/mL fibroblast growth factor bFGF (PeproTech, London, UK). For neurons, cells were seed in poly-L-lysine-coated plates and grown in serum-free Neurobasal medium (Gibco) supplemented with 10 mM L-glutamine, 100 μg/mL penicillin/streptomycin, and 20 μL/mL B-27 supplement. Cells were maintained in a humidified incubator with 5% CO₂ at 37 °C.

Xue Q., Yu C., Wang Y., Liu L., Zhang K., Fang C., Liu F., Bian G., Song B., Yang A., Ju G, & Wang J. (2016). miR-9 and miR-124 synergistically affect regulation of dendritic branching via the AKT/GSK3β pathway by targeting Rap2a. Scientific Reports, 6, 26781.

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Cell Culture Conditions for Various Cell Lines

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J774.1 cells (ATCC; TIB-67) were cultured in DMEM medium supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin, 0.3 mg/ml L-glutamine (Gibco), and 10% FBS (Gibco). A10 cells were purchased at ATCC (CRL-1476) and were cultured in DMEM medium supplemented with 1 mM pyruvate (Gibco), 100 U/ml penicillin, 100 µg/ml streptomycin, 0.3 mg/ml L-glutamine (Gibco) and 10% FBS (Gibco). Vero cells were obtained from WHO (10–87) (originally derived from ATCC (CCL-81)) and were cultured in VP-SFM medium (Gibco), supplemented with 2 mM L-glutamine (Gibco). THP-1 cells used in Figures 1, 2, 3A–C, 4A, Supplementary Figures S1, and S2 were obtained from an in-house stock. To confirm results obtained with this stock, subsequent experiments were performed with THP-1 cells from a stock derived from a vial purchased at ATCC (TIB-202) (Figures 3D, 4B–D, 5, 6, Supplementary Figures S3–S5). THP-1 cells were maintained in RPMI medium (Gibco), supplemented with 50 µM β-mercaptoethanol, 100 U/ml penicillin, 100 µg/ml streptomycin, 0.3 mg/ml L-glutamine (Gibco) and 10% FBS (Gibco). All cell lines were cultivated at 37°C in a humidified atmosphere of 5% CO₂.

Hoonakker M., Zariri A., de Brouwer L., David D., Borgman A, & Sloots A. (2024). An in vitro assay for toxicity testing of Clostridium perfringens type C β-toxin. Frontiers in Immunology, 15, 1373411.

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Cell Culture Conditions for Diverse Cell Lines

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Primary mouse T cells were cultured in T cell media (TCM) that consisted of IMDM supplemented with 10% FBS (HyClone), 50 µM 2-ME (Sigma-Aldrich), 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Gibco). Jurkat and Jurkat-E cells were cultured in RPMI 1640 supplemented with 10% FBS (HyClone), 50 µM 2-ME, 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Gibco). HEK293T were cultured in DMEM supplemented with 10% FBS, 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Gibco). 32D cells were cultured in IMDM supplemented with 10% FBS (HyClone), 10% WEHI-conditioned media (Lee et al., 1982 (link)), 1% L-glutamine (Gibco), and 1% penicillin/streptomycin (Gibco). All cells were cultured at 37°C and 5% CO₂.

Rome K.S., Stein S.J., Kurachi M., Petrovic J., Schwartz G.W., Mack E.A., Uljon S., Wu W.W., DeHart A.G., McClory S.E., Xu L., Gimotty P.A., Blacklow S.C., Faryabi R.B., Wherry E.J., Jordan M.S, & Pear W.S. (2020). Trib1 regulates T cell differentiation during chronic infection by restraining the effector program. The Journal of Experimental Medicine, 217(5), e20190888.

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Cell Culture Protocols for Diverse Cell Lines

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Hap1 TP53 cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) and supplemented with 10% Fetalgro and 5% penicillin and streptomycin (Gibco) and L-glutamine (Gibco). Flp-In T-REx 293 cells AUF1 cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) and supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 5% minimum essential medium nonessential amino acids (100×, Gibco), 5% penicillin and streptomycin (Gibco), and L-glutamine (Gibco).
Additionally, 5 μg mL⁻¹ of blasticidin and 100 μg mL⁻¹ of Zeocin were added to the medium for cell line maintenance. Flp-In T-REx 293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco) and supplemented with 10% Fetalgro bovine growth serum (BGS), 5% minimum essential medium nonessential amino acids (100×, Gibco), 5% penicillin, streptomycin (Gibco), and L-glutamine (Gibco). Additionally, 5 μg mL⁻¹ of blasticidin and 100 μg mL⁻¹ of Zeocin were added to the medium for cell line maintenance. CHO-K cells were cultured in F-12K medium (Kaighn’s modification of Ham’s F-12 medium, Gibco) supplemented with 10% Fetalgro BGS, 5% minimum essential medium nonessential amino acids (100×, Gibco), 5% penicillin and streptomycin (Gibco), and L-glutamine (Gibco).

Powell G., Pavlovic Djuranovic S, & Djuranovic S. (2021). Gene dosage effects of poly(A) track-engineered hypomorphs. Molecular Therapy. Nucleic Acids, 26, 865-878.

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Cell Culture Protocols for Cell Lines

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Hap1 TP53 cells were cultured in IMDM media and supplemented with 10% Fetal-GRO and 5% penicillin and streptomycin (Gibco) and L-glutamine (Gibco). Flp-In T-REx 293 cells AUF1 cell line was cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco) and supplemented with 10% Heat-Inactivated Fetal Bovine Serum (Gibco), 5% minimum essential medium nonessential amino acids (100 × , Gibco), 5% penicillin and streptomycin (Gibco) and Lglutamine (Gibco).
Additionally, 5 μg ml-1 of blasticidin and 100 μg ml-1 of Zeocin was added to media for cell line maintenance. Flp-In T-REx 293 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco) and supplemented with 10% Fetalgro® Bovine Growth Serum (BGS), 5% minimum essential medium nonessential amino acids (100 × , Gibco), 5% penicillin, streptomycin (Gibco), and L-glutamine (Gibco). Additionally, 5 μg ml-1 of blasticidin and 100 μg ml-1 of Zeocin was added to the media for cell line maintenance. CHO-K cells were cultured in F-12K Medium (Kaighn's Modification of Ham's F-12 Medium) (Gibco) supplemented with 10% Fetalgro® Bovine Growth Serum (BGS), 5% minimum essential medium nonessential amino acids (100 × , Gibco), 5% penicillin and streptomycin (Gibco), and L-glutamine (Gibco).

Powell G., Pavlovic‐Djuranovic S., & Djuranović S. (2021). Gene dosage effects of polyA track engineered hypomorphs.

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Cell Line Culture Conditions Standardized

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All the cell lines were cultured as per American Type Culture Collection (ATCC, Manassas, Virgina) recommendations. RL, Raji, MOLM13, U937, were cultured in RPMI (Gibco, Invitrogen, Merelbeke, Belgium), supplemented with 10% fetal calf serum (FSC, Gibco, Invitrogen), 2 mM L-glutamine (Gibco, Invitrogen), 100 IU/mL penicillin (Gibco, invitrogen) and 100 IU/mL streptomycin (Gibco, Invitrogen). Jurkat and HL60 were cultured in IMDM (Gibco, Invitrogen) supplemented with 10 % fetal calf serum (FSC, Gibco, Invitrogen), 2 mM l-glutamine (Gibco, Invitrogen), 100 IU/mL penicillin (Gibco, invitrogen) and 100 IU/mL streptomycin (Gibco, Invitrogen) (complete IMDM, cIMDM). SKOV3 was cultured in DMEM (Gibco, Invitrogen) supplemented with 10% fetal calf serum (FSC, Gibco, Invitrogen), 2 mM L-glutamine (Gibco, Invitrogen), 100 IU/mL penicillin (Gibco, invitrogen) and 100 IU/mL streptomycin (Gibco, Invitrogen). Thp1 was cultured in RPMI (Gibco, Invitrogen) supplemented with 0.05 mM β-mercaptoethanol, 10% fetal calf serum (FSC, Gibco, Invitrogen), 2mM L-glutamine (Gibco, Invitrogen), 100 IU/mL penicillin (Gibco, invitrogen) and 100 IU/mL streptomycin (Gibco, Invitrogen).

De Munter S., Van Parys A., Bral L., Ingels J., Goetgeluk G., Bonte S., Pille M., Billiet L., Weening K., Verhee A., Van der Heyden J., Taghon T., Leclercq G., Kerre T., Tavernier J, & Vandekerckhove B. (2020). Rapid and Effective Generation of Nanobody Based CARs using PCR and Gibson Assembly. International Journal of Molecular Sciences, 21(3), 883.

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Isolation and Culture of Human Mesenchymal Stem Cells

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Mesenchymal stem cells (MSCs) were isolated from human bone marrow (Royal Free, UCL). MSCs adipogenic and osteogenic differentiation capacities were confirmed as described in (39 (link)). MSCs were cultured in αMEM supplemented with 20% Fetal Bovine Serum (FBS), 2 mM L-glutamine and streptomycin/penicillin (Invitrogen). A549 were cultured in F12 supplemented with 10% Fetal Bovine Serum (FBS), 2mM L-glutamine and streptomycin/penicillin (Invitrogen). MDAMB231, H376, A431 and U87MG cells were cultured in DMEM supplemented with 10% Fetal Bovine Serum (FBS), 2mM L-glutamine and streptomycin/penicillin (Invitrogen). Jurkat cells were cultured in RPMI supplemented with 10% Fetal Bovine Serum (FBS), 2mM L-glutamine and streptomycin/penicillin (Invitrogen).

Lourenco S., Teixeira V.H., Kalber T., Jose R.J., Floto R.A, & Janes S.M. (2015). Macrophage Migration Inhibitory Factor - CXCR4 is the dominant chemotactic axis in human mesenchymal stem cell recruitment to tumors. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3463-3474.

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Cell Culture Protocols for Various Cell Lines

Cited 2 times

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GSC lines 387, CW468, D456, and 1517^{68 (link)–70 (link)} were cultured in phenol red-free Neurobasal media plus B27 supplement (Invitrogen), supplemented with 2 mM l-glutamine (Invitrogen), 1 mM sodium pyruvate (Invitrogen), 10 ng/ml basic fibroblast growth factor, 10 ng/ml epidermal growth factor (EGF) (R&D Systems). Cal27, Detroit568 and UMSCC17B were cultured in Dulbecco’s Modified Eagle Medium (DMEM)-F12 1:1 (Gibco), 10% fetal bovine serum (FBS), supplemented with 2 mM l-glutamine (Invitrogen), 0.1 mM MEM non-essential amino acids (NEAA) (Invitrogen) (HNSCC complete media). Cal27-WT and KO cells were cultured in serum-free media (SFM) Defined Keratinocyte SFM (Invitrogen) supplemented with 2 mM l-glutamine (Invitrogen), 0.1 mM NEAA (Invitrogen) and 2 ng/ml EGF (R&D Systems) (Cal27 SFM). HEK 293FT cells were cultured in DMEM (Invitrogen), 10% FBS supplemented by 0.1 mM NEAA, 2 mM l-glutamine, 1 mM MEM sodium pyruvate, 1% PenStrep and 500 μg/ml Geneticin (Invitrogen).

Bernareggi D., Xie Q., Prager B.C., Yun J., Cruz L.S., Pham T.V., Kim W., Lee X., Coffey M., Zalfa C., Azmoon P., Zhu H., Tamayo P., Rich J.N, & Kaufman D.S. (2022). CHMP2A regulates tumor sensitivity to natural killer cell-mediated cytotoxicity. Nature Communications, 13, 1899.

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