The patient-derived GSCs 84NS (RRID: CVCL_C6J0) and 528NS (RRID: CVCL_C6IV) were kindly provided by Dr. Ichiro Nakano (University of Alabama, Birmingham, AL, USA). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM)/F12 (GE Healthcare, Chicago, IL, USA) supplemented with 0.2% B27 (Invitrogen, Carlsbad, CA, USA), 20 ng/mL epidermal growth factor (EGF) (R&D Systems, Minneapolis, MN, USA), 20 ng/mL basic fibroblast growth factor (R&D Systems), 1% penicillin/streptomycin (GE Healthcare), 2 mmol/L L-glutamine (GE Healthcare), and 50 μg/mL gentamicin (Mediatech, Manassas, VA, USA). HEK293T (RRID: CVCL_0063), LN18 (RRID: CVCL_0392), A1207 (RRID: CVCL_8481), and A172 (RRID: CVCL_0131) cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA). These cell lines were cultured in DMEM (GE Healthcare) supplemented with 10% fetal bovine serum (GE Healthcare), 1% penicillin/streptomycin, 2 mmol/L L-glutamine, and 50 μg/mL gentamicin. MG132, CQ, VER-155008, and CHX were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ganetespib was purchased from Abmole Bioscience (Houston, TX, USA). Cell lines used in this study were regularly monitored to ensure the mycoplasma-free cells. In addition, all human cell lines were authenticated in 2019 using short tandem repeat profiling.
L glutamine
Manufactured by GE Healthcare
Sourced in Austria, United States, Germany, United Kingdom, Italy, Switzerland, France, Belgium
L-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 metabolism of glucose and other nutrients. L-glutamine is an important component in cell culture media and is commonly used in laboratory settings for the growth and maintenance of cells.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by GE Healthcare (45 mentions)
Penicillin, by GE Healthcare (44 mentions)
Penicillin streptomycin, by GE Healthcare (35 mentions)
Fetal bovine serum (fbs), by GE Healthcare (25 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (23 mentions)
Rpmi 1640, by GE Healthcare (17 mentions)
Dulbecco modified eagle medium (dmem), by GE Healthcare (17 mentions)
Penicillin, by Thermo Fisher Scientific (17 mentions)
Streptomycin, by Thermo Fisher Scientific (17 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (16 mentions)
Fetal bovine serum (fbs), by Merck Group (14 mentions)
L glutamine, by Thermo Fisher Scientific (14 mentions)
Fetal calf serum (fcs), by GE Healthcare (12 mentions)
Penicillin, by Corning (11 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (11 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (11 mentions)
Streptomycin, by Corning (10 mentions)
Rpmi 1640 medium, by Merck Group (9 mentions)
Trypsin edta, by GE Healthcare (9 mentions)
Fetal bovine serum (fbs), by Lonza (8 mentions)
232 protocols using l glutamine
1
Cultivation and Authentication of Human Glioma Cell Lines
2
Establishment of GBM Cell Lines
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Human U343MG cells were obtained from the American Type Culture Collection (ATCC) and cultured in Basal Medium Eagle (Thermo Fisher Scientific), supplemented with 10% fetal calf serum, 10 mM HEPES, 2 mM L-glutamine, and 1% non-essential amino acids (all from GE Healthcare) at 37°C in a humidified atmosphere containing 5% CO2. The human DD-T4 cell line was grown in Dulbecco's Modified Eagle's Medium (plus GlutaMAX-I) supplemented with 10% fetal calf serum and 1% non-essential amino acids (all from GE Healthcare) at 37°C and 8.5% CO2. Human GBM stem-like cells GS-8 [53 (link)] were kindly provided by K. Lamszus (University Medical Center Hamburg-Eppendorf, Germany) via material transfer agreement and grown in stem cell medium: Neurobasal Medium (Thermo Fisher Scientific) supplemented with 2 mM L-glutamine (GE Healthcare), 32 U/ml Heparin (Merck Millipore), B27 supplement, 20 ng/ml human EGF and 20 ng/ml human FGFb (all from Thermo Fisher Scientific) at 37°C and 5% CO2. To generate GS-8_GFP/fLuc cells, lentiviral transduction of GS-8 cells was kindly performed as described [54 (link)] by M. Cartellieri (Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Germany) using the lentiviral vector p6NST50 [55 (link)] containing the firefly luciferase gene.
3
Isolation and Enrichment of B Cell Subsets
4
A2780 Cell Culture with DMEM
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DMEM cell culture media, penicillin/streptomycin mixture, foetal bovine serum, l -glutamine, and trypsin were all purchased from PAA Laboratories GmbH. DMEM was supplemented with foetal bovine serum (50 mL), penicillin/streptomycin mixture (5 mL), and l -glutamine (5 mL). The A2780 cell line was purchased from the European Collection of Cell Cultures.
5
Generation of Stable EGFP-Expressing Cell Lines
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A human chronic myelogenous leukaemia cell line (K562) stably expressing an EGFP gene from the AAVS1 locus was prepared by nucleofection of K562 cells (ATCC) with the pAAVS1_SA2APuro_PGK_EGFP_HSVtkpA plasmid (created by cloning PGK-EGFP-HSVtkpA cassette into Addgene plasmid #22075) along with AAVS1-specific ZFN expression plasmids19 (link). After initial selection with 0.5 μg/ml of puromycin (Invitrogen), single cell clones were analysed by PCR and a clone containing a single targeted integration of the EGFP expression cassette was chosen for further experiments. K562-EGFP cells were cultured in RPMI medium (Gibco/Life Technologies) supplemented with 10% FCS (Gibco/Life Technologies), 100 IU penicillin and 10 mg/ml streptomycin (P/S; PAA Laboratories GmbH, Austria), and 2 mM L-glutamine (PAA Laboratories). The murine embryonic stem cell line BK4-G3, containing a single copy of the EGFP gene in the HPRT locus, has previously been described20 (link). Cells were grown on gamma-irradiated mouse embryonic fibroblasts (MEFs) in knockout DMEM (Gibco/Life Technologies) supplemented with 15% FCS (PAN Biotech, Berlin, Germany), leukaemia inhibitory factor (LIF)-conditioned medium produced from recombinant Chinese hamster ovary cells (diluted 1:500), P/S, 2 mM L-glutamine, 0.1 mM non-essential amino acids (PAA Laboratories), and 0.1 mM β-mercaptoethanol (Gibco/Life Technologies).
6
Murine Skeletal Myoblast Differentiation
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The murine skeletal myoblast cell line PMI28 [18 ] was cultured in a growth medium composed of Ham’s F10 (PAA Laboratories GmbH, Pasching, Austria), supplemented with 20% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA), 2 mM L-glutamine (PAA Laboratories), and Penicillin (100 I.U./ml) / Streptomycin (100 μg/ml, PAA Laboratories). 24 hours after seeding of the cells the growth medium was replaced by a differentiation medium containing DMEM medium with 2% horse serum (Gibco, Life Technologies GmbH, Darmstadt, Germany), 2 mM L-glutamine (PAA Laboratories), and Penicillin (100 I.U./ml) / Streptomycin (100 μg/ml) (PAA Laboratories). The differentiation medium of the treatment groups additionally contained 2 x 103 U/ml murine recombinant TNF-α (Roche Diagnostics, Rotkreuz, Switzerland) or 5 ng/ml murine recombinant IGF1 (Sigma-Aldrich). The control and treatment media were replenished twice a day to ensure cytokine and growth factor activity. Murine PMI28 cells were harvested 24 h after the induction of fusion by serum withdrawal for RNA analyses. Cells were propagated and differentiated at 37°C in 80% relative humidity and 5% CO2.
7
Isolation and Enrichment of B Cell Subsets
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HEK293 cells were grown in suspension at 37 °C in Pro-293 medium (Lonza) supplemented with 1.5% fetal bovine serum (Biosera) and 2 mM l-glutamine (PAA Laboratories). Primary splenocytes were obtained from the spleen by passing through a 70 μm cell strainer and lysing red blood cells using ACK buffer (Gibco). Naive B cells were isolated by negative selection using anti-CD43 microbeads (Miltenyi Biotech). GC B cells were enriched from splenocytes of immunized mice by negative selection using biotinylated anti-CD43, anti-IgD and anti-CD11c antibodies and anti-biotin microbeads (Miltenyi Biotech). For the enrichment of transferred B1-8 B cells, the negative selection was supplemented with biotinylated anti-CD45.1, which is expressed by the host cells. After enrichment, cells were recovered in RPMI 1640 media (Sigma) supplemented with 10% fetal bovine serum (Biosera), 1% MEM non-essential amino acids (GIBCO), 2 mM l-glutamine (PAA Laboratories), 50 μM 2- mercaptoethanol (GIBCO), 100 U/ml penicillin and 100 μg/ml streptomycin.
8
Bronchial Epithelial Cell Culture and Analysis
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Bronchial epithelial cell growth medium (BEGM) with antibiotics was purchased from Lonza (Walkersville, MD). The BEGM was prepared following manufacturer’s guideline, which contained all the supplements (BPE, hEGF, epinephrine, transferrin, insulin, retinoic acid, triiodothyronine, GA) except hydrocortisone to avoid any inhibitory effect of corticosteroids on cell pro-inflammatory responses. RNA lysis buffer RLT was from Qiagen (Hilden, Germany). RIPA western lysis buffer was purchased from Thermo-Fisher Scientific (Waltham, MA). DMEM (high glucose) for making D10 (DMEM + 10% FBS + 1% Pen/Strep + 1% Amphotericin B + 1% L-Glutamine+ 0.5% Gentamicin) was from GE Life Sciences (Logan, UT). The nuclear extraction kit and TransAM NF-κB p65 assay kit were from Active Motif (Carlsbad, CA). IL-8, IP-10 and TNF-α ELISA kits were obtained from R&D systems (Minnieaplois, MN).
9
Lipid Nanoparticle Preparation and Characterization
10
Culturing Four AML Cell Lines
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Four AML cell lines, MOLM-13 (AML M5), KASUMI-1 (AML M2), OCI-AML3 (AML M4) and NOMO-1 (AML M5) were obtained from the American Type Culture Collection, and were mycoplasma-tested and authenticated using the LGC Standards Cell Line Authentication service. The cell lines were cultured at 37°C in a 5% CO2 atmosphere at a density of 0.3×106 cells/ml in complete medium, in T75 flasks. MOLM-13 and KASUMI-1 cells were cultured in RPMI-1640 (Euroclone) supplemented with 20% heat-inactivated FBS (GE Healthcare), 2 mM L-glutamine (GE Healthcare), 100 U/ml penicillin, 100 µg/ml streptomycin (GE Healthcare) and 0.2% Mycozap (Lonza, Inc.). OCI-AML3 cells were cultured in α-MEM (Lonza, Inc.) with 20% FBS, 2 mM L-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin. NOMO-1 cells were grown in RPMI-1640 with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin.
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