Pairs of Rat E18 cortices (Cat. No. SDECX, BrainBits, LLC) were dissociated for 10 min in papain (Cat. No. LK003176; Worthington Biochemical; at least 20 U/ml). A single-cell suspension was obtained and filtered through a 40 μm cell strainer. The resulting primary cells were cultured in NeuroCult™ Neuronal Plating medium (Cat. No. 05713, STEMCELL Technologies) or Neurobasal medium for Neurobasal and NEUMO cultures (Cat. No. 21103-049, Thermo Fisher Scientific) with 1× SM1 (Cat. No. 05711, STEMCELL Technologies), 0.5 mM l -glutamine (Cat. No. 07100, STEMCELL Technologies) and 25 μM l -glutamic acid (Cat. No. G8415, Sigma) on culture-ware pre-coated with 10 µg/ml poly-D-lysine (Cat. No. P7280, Sigma). Cells were plated at 30,000 cells/cm2 in a 24-well plate. Five days post-plating, half media changes were performed every 3–4 days with either BrainPhys™ (Cat. No. 05790, STEMCELL Technologies), Neurobasal, BrightCell™ NEUMO (Cat. No. SCM146, Merck) BrainPhys™ without Phenol Red (Cat. No. 05791, STEMCELL Technologies), or BrainPhys™ Imaging (Cat. No. 05796, STEMCELL Technologies) supplemented with 1× SM1. Neurobasal and BrightCell™ NEUMO media were also supplemented with 0.5 mM l -glutamine.
L glutamine
Manufactured by STEMCELL
Sourced in Germany, United States, Canada
L-glutamine is a common amino acid supplement used in cell culture media. It serves as a primary energy source for cells and supports various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by STEMCELL (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by STEMCELL (4 mentions)
Gentamycin, by Merck Group (3 mentions)
Hank s basal salt solution, by STEMCELL (3 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (3 mentions)
Trypsin, by Thermo Fisher Scientific (3 mentions)
Erythropoietin, by STEMCELL (3 mentions)
Poly l lysine (pll), by Merck Group (3 mentions)
Interleukin 2 (il 2), by Thermo Fisher Scientific (3 mentions)
Non essential amino acids (neaa), by STEMCELL (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Dmem f12, by Thermo Fisher Scientific (3 mentions)
Hepes, by Thermo Fisher Scientific (2 mentions)
Anti mouse cd28, by Thermo Fisher Scientific (2 mentions)
Complete rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Anti mouse cd3, by Thermo Fisher Scientific (2 mentions)
β mercaptoethanol, by Maclin Biochemical Technology (2 mentions)
Adenosine, by Merck Group (2 mentions)
46 protocols using l glutamine
1
Rat E18 Cortical Neuron Culture
2
CHO Cell Culture Protocol
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Chinese Hamster Ovary cells (European Collection of Authenticated Cell Cultures ECACC, cells CHO-K1, cat. no. 85051005, obtained directly from the repository) were grown in 25 cm2 culture flasks (TPP, Trasadingen, Switzerland) in HAM F-12 growth medium (PAA, Kremplstraße, Austria) for 2–3 days in an incubator (Kambič, Semič, Slovenia) at 37 °C and humidified 5% CO2. The growth medium (used in this composition through all experiments) was supplemented with 10% fetal bovine serum (Sigma-Aldrich, Darmstadt, Germany), L-glutamine (StemCell, Vancouver, CAN) and antibiotics penicillin/streptomycin (PAA) and gentamycin (Sigma-Aldrich, Germany). On the day of the experiments, the cell suspension was prepared. Cells were detached by 10× trypsin-EDTA (PAA), diluted 1:9 in Hank’s basal salt solution (StemCell) and the trypsin was inactivated by 2.5 ml of the HAM F-12 growth medium. Cells were transferred to a 50 ml centrifuge tube (TPP) and centrifuged 5 min at 180 g and 22 °C. The supernatant was removed, and cells were re-suspended in the growth medium HAM F-12 at cell density 107 cells/ml.
3
Culturing M3-9-M RMS Cells
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M3‐9‐M RMS cells (gift from Mackall Lab, Stanford) were cultured in RPMI media 1640 (Cat# 350‐000‐CL, Wisent Inc) containing 1% l ‐glutamine (Cat# 07100, STEMCELL Technologies), 10% heat‐inactivated foetal bovine serum (Cat# 12483‐020, Gibco), HEPES (Cat# 15630‐080, Gibco), 1% non‐essential amino acids (Cat# 11140‐050, Gibco), 1% sodium pyruvate (Cat# 11360‐070, Gibco), 1% penicillin/streptomycin (P/S) (Cat# 15140‐122, Thermo Fisher Scientific) and 50 mM 2‐mercaptoethanol (Cat# 21985‐023, Gibco).17
4
Mouse Oocyte Maturation and Fertilization
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Ten‐month‐old mice were used to obtain GV‐ and MII‐stage oocytes. At 48 h after the injection of PMSG (Ningbo, China) into oocytes, GV‐stage oocytes were retrieved from the ovaries of mice and moved to M2 medium (Sigma, m7167) containing 0.2 mM IBMX (Sigma, i5879). After microinjection, the oocytes were washed and moved into IVM medium containing TCM‐199 (Gibco, 12340030) supplemented with 10% FBS (Gibco, 10099141C), 0.2 mmol/L sodium pyruvate (Sigma, P5280), 2 mmol/L L‐glutamine (Stem Cell, 07100), 10 IU/ml PMSG (Ningbo, China), 10 IU/ml HCG (Ningbo, China), 1 μg/ml β‐oestradiol (Sigma, E2758) and 10 ng/ml EGF for culture in a 37°C incubator containing 5% CO2. For MII oocytes, mice were injected with HCG 48 h after PMSG, MII oocytes were retrieved from fallopian tubes 12 h after HCG (Ningbo, China) injection, and cumulus cells were digested by hyaluronidase (1 mg/mL, Sigma, H3506). The fertilized oocytes were transferred to the oviducts of surrogate mothers on an ICR background.
5
Isolation and Culture of Primary PSCs
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The mice were sacrificed and their femurs and tibias were dissected. After removing the epiphysis, the bone was flushed for removing total bone marrow cells. To obtain primary PSCs, the remaining bone-washed explants without muscles and tendons were cultured using Mouse MesenCultTM Expansion Kit (# 05513, STEMCELL Technologies, United States) supplemented with L-Glutamine (# 07100, STEMCELL Technologies); the PSCs migrated from the explants within 3 days. After 2 weeks, the bones were removed, and the PSCs were digested with trypsin and directly used for in vitro and in vivo experiments without further amplification.
6
Generating Engineered Cell Lines for Research
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BON1-SSTR2 cells were generated by transfecting wildtype BON1 cells with the pcDNA3.1-SSTR2 vector as described previously 23 (link), using Lipofectamine Transfection Reagent (ThermoFisher Scientific). Selection was done for 1 week using 2 mg/mL G418 (Invivogen), after which cells were cultured in presence of 500 μg/mL G418 every other passage. BON1-SSTR2 cells were cultured in DMEM/F12 medium (Gibco), supplemented with 10% fetal calf serum (FCS; Biowest) and 1% penicillin-streptomycin (PS; Sigma-Aldrich). GOT1 cells were maintained in RPMI 1640 (Gibco), supplemented with 10% FCS, 1% PS, 5 mM L-glutamine (Stemcell Technologies), 5 µg/mL insulin (Sigma) and 5 µg/mL human holo-transferrin (Sigma). NCI-H69 cells (ATCC) were cultured in RPMI 1640, supplemented with 10% FCS and 1% PS. All cells were cultured at 37 °C and 5% CO2.
7
Naive CD4+ T Cell Differentiation
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Naïve CD4+ T cells were sorted from wild-type C57BL/6 mouse spleens using an EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit (STEMCELL). About 500,000 cells were plated in 48-well plates in 0.5 mL of complete RPMI 1640 medium (Gibco) supplemented with 10% FBS (Gibco) and stimulated with anti-mouse CD3 (5 μg/mL, eBioscience) and anti-mouse CD28 (2 μg/mL, eBioscience) plus IL-2 (10 ng/mL, PeproTech), TGF-β1 (50 ng/mL, PeproTech), 2 mM L-glutamine (STEMCELL), and 50 mM β-mercaptoethanol (Macklin) in the absence or presence of 2 μM luteolin (RHAWN™) for 4 d at 37°C with 5% CO2. Finally, CD4-FITC, CD25-PE, and FOXP3-APC were used to analyze the cells.
8
Culturing L. donovani Promastigotes
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L. donovani Sudan strain S2 promastigotes were routinely cultured in M199 (Sigma-Aldrich) with 10% heat inactivated fetal bovine serum (FBS, Gibco), 20 mM HEPES (Sigma-Aldrich), 6 μg/mL hemin (Sigma-Aldrich), 10 μg/mL folic acid (Sigma-Aldrich), 2 mM L-glutamine (Stemcell), 100 U/mL penicillin/streptomycin (Sigma-Aldrich) and 100 μM adenosine (Sigma-Aldrich) at 26°C. Every 3 days the parasites were subcultured 1:10 in fresh medium and were kept in culture for a maximum of 20–25 passages. In order to maintain virulence and infectivity, fresh parasites were routinely obtained by purification of amastigotes from spleens of 6–8 week infected Syrian Golden hamsters followed by in vitro transformation into promastigotes by culturing for 5–7 days at 26°C in promastigote medium. Prior approval for animal experiments was obtained from the Animal Care Committee of University of British Columbia (approval# A14-00218)
9
Maintaining and Differentiating Mouse Embryonic Stem Cells
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Mouse ES cells (E14TG2a) were maintained on gelatin-coated dishes in ATCC-formulated Dulbecco’s Modified Eagle’s Medium (ATCC), supplemented with 0.1 mM of 2-mercaptoethanol, 10% fetal bovine serum (StemCell Technologies), 2 mM L-glutamine, 0.1 mM MEM nonessential amino acid, 100 U/mL penicillin, 10 µg/mlstreptomycin, and 10 ng/mL LIF (StemCell Technologies). The mES cells were passaged every 2 d at a ratio of 1:5 by washing with PBS, dissociating with 0.25% trypsin (GIBCO) for 3 min at 37°C, and resuspending in mES media. Media was changed daily. To induce differentiation, the mES cells were passaged and then cultured in ES cell culturing media without LIF. Media was changed every 2 d. The undifferentiated (E14-d0) and differentiating (E14-d6) states of mES cells were verified by SSEA-1 (stage-specific embryonic antigen-1) staining with StainAlive SSEA-1 Antibody (DyLight 488) (Stemgent) and quantitative RT-PCR analysis of relative expression levels of three major pluripotency factors including Nanog, Sox2, and Pou5f1.
10
Isolation and Expansion of Murine Bone Marrow Mesenchymal Stem Cells
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BM cells were extracted from wild-type or GFP mice by flushing the marrow from femurs and tibias into cold sterile DMEM/F12 (ThermoFisher Scientific, Waltham, CA) medium. Cells were filtered through a 70 µM filter and centrifuged at 700 × g for 8 min at 4°C. Cells (5 × 107) were plated in a T75 flask in 15 ml murine MSC-specific expansion medium (Mouse MesenCult Expansion Kit, Stem Cell Technologies, Cambridge, MA) containing l -Glutamine 1% (Stem Cell Technologies) and antibiotic/antimycotic solution 1% (Sigma Aldrich, St. Louis, MO). Cells are cultured at 37°C with 5% CO2 and atmospheric O2 concentration (∼20%). At the second passage, MSCs were collected and used for BM programing experiments.
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