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Dulbecco s modified eagle medium

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Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium that provides essential nutrients for the growth and maintenance of various cell types. It is a widely used basal medium in the field of cell biology and biotechnology.

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Lab products found in correlation

Fetal bovine serum (fbs), by Merck Group (99 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (80 mentions) Streptomycin, by Merck Group (54 mentions) Penicillin, by Merck Group (53 mentions) Penicillin streptomycin, by Merck Group (46 mentions) Streptomycin, by Thermo Fisher Scientific (42 mentions) Penicillin, by Thermo Fisher Scientific (42 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (34 mentions) L glutamine, by Merck Group (28 mentions) Dimethyl sulfoxide (dmso), by Merck Group (27 mentions) L glutamine, by Thermo Fisher Scientific (22 mentions) Phosphate buffered saline (pbs), by Merck Group (18 mentions) Rpmi 1640, by Merck Group (16 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (15 mentions) Bovine serum albumin (bsa), by Merck Group (13 mentions) Rpmi 1640 medium, by Merck Group (12 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (11 mentions) Glutamine, by Merck Group (10 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (10 mentions) Mcf 7, by American Type Culture Collection (10 mentions)

499 protocols using dulbecco s modified eagle medium

1

GT Cytotoxicity in Colon Cancer Cells

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HCT116 (p53−/−) and HCT116 (p21−/−) cells were grown in Dulbecco's Modified Eagle Medium (Sigma−Aldrich, Munich, German) supplemented with sodium pyruvate and 4500 mg/L glucose. Roswell Park Memorial Institute (RPMI) 1640 with 25 mm N-2-hydroxyethylpiperazine-N-ethanesulfonic acid (HEPES) and L-glutamine was used as a medium for HCT116 (p53+/+) human colon cancer cells. One percent penicillin-streptomycin (100 U/mL) and 10% heat-inactivated foetal bovine serum (FBS) were added to all cell media. All culture cells were grown in a humidified atmosphere of 5% carbon dioxide and 95% air.
A stock of 10 mg GT (Sigma-Aldrich) was dissolved in 50 µL 70% ice-cold ethanol and, depending on the cell type, 950 µL either Dulbecco's Modified Eagle Medium or RPMI. The cells were treated with different drug concentrations (0–60 µg/mL). Depending on the experiment done, the 3 cell lines were plated in 100 mm, 60 mm, 6-well, or 96-well plates at a plating density of 105 for senescence and proliferation analyses or 1.2  × 105 cells/mL for proteins. At 50% confluency, the cells were treated and incubated with GT for 6, 15, 24, 48, or 72 hours. The HCT116 p53 null cells were kindly provided by Dr Carlos Galmarini, MD, PhD (INSERM, Lyon, France) and the HCT116 p53+/+ and p21−/- were provided by Dr Regine Schneider-Stock, PhD (Erlangen, Germany).
Houssein M., Abi Saab W., Khalil M., Khalife H, & Fatfat M. (2020). Cell Death by Gallotannin Is Associated with Inhibition of the JAK/STAT Pathway in Human Colon Cancer Cells. Current Therapeutic Research, Clinical and Experimental, 92, 100589.
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2

Cell Culture of Breast Cancer Cell Lines

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The MCF-7 cells were cultured in RPMI-1640 (Sigma, USA) while MDA-MB-231 was maintained in Dulbecco’s modified eagle medium (Sigma, USA). The MCF-10a cells were cultured in the mixture medium of Ham’s F-12 (Sigma, USA) and Dulbecco’s modified eagle medium supplemented with 20 ng/mL epidermal growth factor (EGF), 10 μg/mL insulin, and 250 ng/μL hydrocortisone. All media were added with 10% (v/v) heat-inactivated foetal bovine serum (FBS) (PAA, Austria), penicillin (100 I.U/mL) and streptomycin (100 ng/mL) (PAA, Austria). The cells were cultured at 37 °C in a 90% humidified incubator with 5% CO2. When the cells were 80% confluent, they were sub-cultured to a fresh media.
Razak N.A., Abu N., Ho W.Y., Zamberi N.R., Tan S.W., Alitheen N.B., Long K, & Yeap S.K. (2019). Cytotoxicity of eupatorin in MCF-7 and MDA-MB-231 human breast cancer cells via cell cycle arrest, anti-angiogenesis and induction of apoptosis. Scientific Reports, 9, 1514.
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3

Matrigel Invasion Assay for Cell Migration

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Cells were transfected in 60 mm tissue culture plates for 24 hours. Harvested cells were counted on the Z2 Coulter Counter Analyzer (Beckman Coulter). Cells were then resuspended in Dulbecco's Modified Eagle Medium (Sigma-Aldrich) without serum. The resuspended cells were then pipetted into a Corning BioCoat Matrigel Invasion Chamber (ThermoFisher), 8 μm pore size, in 24-well tissue culture-grade plates. Dulbecco's Modified Eagle Medium (Sigma-Aldrich) supplemented with 10% FBS (Atlas Biologicals) was placed in the well of the plate as a chemo-attractant. After 24 hours, non-invading cells were removed using a cotton-tipped swab. Invasive cells, embedded into the Matrigel, was fixed using 70% ethanol then stained with a 0.5% crystal violet solution. Microscopy was performed with the Evos XL Cell Imaging System (ThermoFisher). Cells were counted in five fields of view at 10× magnification and then averaged as cells per 10× field.
Vo D.T., Karanam N.K., Ding L., Saha D., Yordy J.S., Giri U., Heymach J.V, & Story M.D. (2019). miR-125a-5p Functions as Tumor Suppressor microRNA And Is a Marker of Locoregional Recurrence And Poor prognosis in Head And Neck Cancer. Neoplasia (New York, N.Y.), 21(9), 849-862.
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4

Generation of HIV-1 Viral Stocks in 293T Cells

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Viral stocks of HIV-1JR-CSF were generated by transfecting HIV-1JR-CSF DNA50 (link)–54 (link) into 293T cells (American Tissue Culture Collection, catalog number CRL-3216, passage <10) using Lipofectamine 2000 (Invitrogen). 293T cells were cultured at 37 °C, 10% CO2 in Dulbecco’s Modified Eagle Medium (Sigma) supplemented with 10% fetal bovine serum, 25 mM HEPES, 500 units/ml penicillin, 50 µg/ml streptomycin and 2 mM L-glutamine (Cellgro), cells were regularly checked for morphology by microscopy. Viral particles were collected in tissue culture medium that was centrifugated at 2000×g for 20 min at 4 °C to remove cell debris. Tissue culture infectious units (TCID)/ml of HIV were determined by titration using TZM-bl cells (NIH AIDS Research and Reference Reagent Program, catalog number 8129, passage number 2–6). Infected TZM-bl cells were stained using a solution containing 4 µM potassium ferrocyanide, 4 µM potassium ferricyanide, 2 µM magnesium chloride, 0.4 mg/ml X-gal55 (link),56 (link) and stained cells counted. TZM-bl cells were cultured at 37 °C 10% CO2 in TZM-bl medium (Dulbecco’s Modified Eagle Medium (Sigma) supplemented with 10% heat-inactivated fetal bovine serum, 25 mM HEPES, 500 units/ml penicillin, 50 µg/ml streptomycin and 2 mM L-glutamine (Cellgro)), and regularly checked for morphology by microscope.
Benhabbour S.R., Kovarova M., Jones C., Copeland D.J., Shrivastava R., Swanson M.D., Sykes C., Ho P.T., Cottrell M.L., Sridharan A., Fix S.M., Thayer O., Long J.M., Hazuda D.J., Dayton P.A., Mumper R.J., Kashuba A.D, & Victor Garcia J. (2019). Ultra-long-acting tunable biodegradable and removable controlled release implants for drug delivery. Nature Communications, 10, 4324.
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5

Protein Expression and Extraction from HEK293T and N2A Cells

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Cells were maintained in a 37 °C incubator with 5% CO2. HEK293T cells were cultured in Dulbecco’s Modified Eagle Medium (Sigma) supplemented with 10% foetal bovine serum (FBS) (Gibco) and 5 U ml− 1 Penstrep (Lonza). Neuro-2a(N2A) (ATCC) cells were cultured in Dulbecco’s Modified Eagle Medium (Sigma) supplemented with 10% FBS (Gibco), 5 U ml− 1 Penstrep (Lonza) and 5 mM sodium pyruvate.
HEK293T and N2A cells were transfected with 700 ng of plasmid using 3.5 μg PEI/ml media and one tenth media volume of OptiMEM in a 24 well format. Approximately, 50,000 HEK293T cells were seeded / well and 75,000 N2A cells were seeded per well of the 24 well plate. Proteins were extracted 72 h post-transfection. Cells were washed in ice cold phosphate buffered saline (PBS) and subsequently lysed in ice cold lysis buffer (50 mM Hepes pH 7.5, 150 mM NaCl, 10% glycerol, 0.5% Triton X-100, 1 mM EDTA, 1 mM DTT, protease inhibitor cocktail (Sigma)) for 10 min on ice. Extracts were then centrifuged at 17,000 g for 5 min at 4 °C. Extracts were quantified using Bradford Reagent (BioRAD), resolved by SDS-PAGE, electroblotted onto nitrocellulose membrane and probed to the relevant primary antibodies.
Shaw M.P., Higginbottom A., McGown A., Castelli L.M., James E., Hautbergue G.M., Shaw P.J, & Ramesh T.M. (2018). Stable transgenic C9orf72 zebrafish model key aspects of the ALS/FTD phenotype and reveal novel pathological features. Acta Neuropathologica Communications, 6, 125.
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6

Cell Culture of Human Cancer and Control Cells

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The cell culture was carried out with standard cell culture methods. Human colorectal adenocarcinoma cells (Caco-2) were accessed from Yeditepe University Department of Medical Genetics Research Group and cultured in a Dulbecco's Modified Eagle Medium (Sigma Aldrich) with 10 % (v/v) Fetal Bovine Serum (BioSera) and 1 % (v/v) penicillin-streptomycin (GenMarkBio) at 37 °C and 5 % CO2. Human vascular endothelial cells (HUVECs) were obtained from Bahc¸es¸ehir University Department of Medical Genetics Research Group and cultured in Dulbecco's Modified Eagle Medium (Sigma Aldrich) with 10 % (v/v) Fetal Bovine Serum and 1% (v/v) penicillin-streptomycin (GenMarkBio) at 37 °C and 5 % CO2. CCD-1072Sk (Human skin fibroblast) cells were procured from Bezmialem Vakif University Department of Medical Genetics Research Group as a control group and cultured in Eagle Minimum Essential Medium (Sigma Aldrich) with 10 % (v/v) Fetal Bovine Serum and 1 % (v/v) penicillinstreptomycin (GeneMarkBio) at 37 °C and 5 % CO2. All cells used in the study were under 25 passages for all experiments.
Kucak M., Demir U, & Muslumanoglu M.H. (2022). Investigation of the cytotoxic, anti-proliferative and anti-angiogenic effects of Toluhydroquinone on Caco-2 cell line. Cellular and molecular biology (Noisy-le-Grand, France), 68(8).
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7

Collagen Scaffold Fabrication with Embedded Cells

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Sterile polydimethylsiloxane (PDMS, SYLGARD™ 184, Dow Corning) wells (10 mm diameter, 1 mm height) were placed in a 24 well plate to ensure uniform collagen scaffold geometry and electric field distribution between each replicate. PDMS wells were treated with 1% PEI (Acros Organics) for 10 minutes, 0.1% glutaraldehyde (Fisher Scientific) for 20 minutes, and then washed twice with deionized water prior to collagen seeding to ensure collagen adhesion during treatment. Commercial rat tail collagen type I (BD Biosciences) was neutralized using a solution of 10X Dulbecco’s Modified Eagle Medium (10% total volume, Sigma Aldrich), 1 N NaOH (2% collagen volume, Sigma Aldrich), and 1X Dulbecco’s Modified Eagle Medium (Sigma Aldrich) to a final concentration of 5 mg/mL. U251 cells were detached from flasks using 0.25% trypsin/EDTA (Thermo Fisher Scientific) solution and added to the neutralized collagen solution at a concentration of 1×106 cells/mL. The collagen/cell solution was dispensed into PDMS wells and PDMS tops were used to mold the collagen flat while they polymerized in a cell culture incubator for 20 minutes. PDMS tops were removed and cell culture media was added. Collagen scaffolds were maintained in the incubator for 24 hours prior to treatment.
Rodermel S., Haley J., Jiang C.Z., Tsai C.H, & Bogorad L. (1996). A mechanism for intergenomic integration: abundance of ribulose bisphosphate carboxylase small-subunit protein influences the translation of the large-subunit mRNA.. Proceedings of the National Academy of Sciences of the United States of America, 93(9), 3881-3885.
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8

Isolation and Culture of Cerebral Microvessels

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Cerebral microvessels were isolated from the brains of male C57BL/6 mice based on a previously reported method (Yin et al., 2006). Cerebral cortices were dissected and homogenized. After centrifugation (10,000 × g for 5 minutes at 4°C), the precipitates were collected and resuspended in PBS. After the second centrifugation, the precipitates were resuspended in 18% dextroside and centrifuged again (1500 × g for 20 minutes at 4°C). To avoid contamination from neurons and glial cells in microvascular tissues, the collected precipitates were resuspended in 18% dextroside and centrifuged (10,000 × g for 1 minute at 4°C). The precipitates were resuspended in PBS and then filtered through a 70-μm membrane. The filtrate was discarded, and the final vessel pellets were collected and stored at –80°C for biochemical assays. Cells in cerebral microvessels were cultured with Dulbecco’s modified Eagle medium (MilliporeSigma) containing 10% fetal bovine serum (Gibco Life Technologies, Grand Island, NY, USA) to confluence. Mouse cerebral vascular endothelial cell cultures were exposed to OGD for various time points (1, 2, 4, 6, 24 and 48 hours) (Yin et al., 2013). The culture medium was replaced with glucose-free Dulbecco’s modified Eagle medium (MilliporeSigma), then cells were incubated at 37°C in 5% CO2. Cells were exposed to OGD for 24 hours.
Wu N., Cheng C.J., Zhong J.J., He J.C., Zhang Z.S., Wang Z.G., Sun X.C, & Liu H. (2022). Essential role of MALAT1 in reducing traumatic brain injury. Neural Regeneration Research, 17(8), 1776-1784.
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9

Synthesis of Gold and Silver Nanoparticles

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Hydrogen tetra-chloroauric hydrate (HAuCl4 xH2O, 99.9%), sodium borohydride (NaBH4, 99%), silver nitrate (AgNO3, 99%), cetyltrimethylammonium bromide (CTAB, ≥99%), Hydroquinone (HQ, 99%), sodium oleate, (NaOL, ≥99%), hydrochloric acid (HCl, (12.1 M)), Dulbecco’s Modified Eagle Medium (DMEM), Dulbecco’s phosphate-buffered saline (PBS), Roswell Park Memorial Institute (RPMI) medium, graphite powder, sulfuric acid (H2SO4, 95.0–98.0%), sodium nitrate (NaNO3, 99%), potassium permanganate (KMnO4, 99%), peroxide (H2O2, 30%), and sodium hydroxide (NaOH, ≥98%) were purchased from Sigma-Aldrich, (Kempton Park, South Africa). All glassware used in the experiments was cleaned, washed thoroughly with MilliQ water (15.0 MΩ cm @ 25 °C), and dried before use.
Lebepe T.C, & Oluwafemi O.S. (2022). Thermal and Medium Stability Study of Polyvidone-Modified Graphene Oxide-Coated Gold Nanorods with High Photothermal Efficiency. Nanomaterials, 12(19), 3382.
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10

Generating Peptide-Resistant Breast Cancer Cells

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MDA-MB-231 breast cancer cells were obtained from Dr. S. Drover (Memorial University of Newfoundland, NL, Canada). NRC-03-and NRC-07-resistant MDA-MB-231 cells were generated by continuous exposure of MDA-MB-231 cells to increasing concentrations of DAA peptides NRC-03 or NRC-07 for approximately one year. Peptide-resistant cells were capable of growing in the presence of 50 μM NRC-03 or NRC-07. However, treatment with DAA peptides NRC-03 or NRC-07 at greater than 50 μM resulted in excessive toxicity, indicating that only low-level resistance was generated. Parental MDA-MB-231 cells cultured for the same amount of time as resistant cells but in the absence of peptide were used as a control for all experiments. All cells were grown in Dulbecco’s Modified Eagle Medium (Sigma-Aldrich Canada, Oakville, ON, Canada) supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine, 5 mM HEPES (pH 7.4), and 2.5% heat-inactivated fetal bovine serum (Invitrogen, Burlington, ON, Canada). Cells were seeded, in peptide-free medium, into tissue culture plates, and were cultured for 24 h to promote cell adhesion. Stock flasks were passaged as required to maintain optimal cell growth and were routinely confirmed to be free of mycoplasma contamination.
Hilchie A.L., Gill E.E., Coombs M.R., Falsafi R., Hancock R.E, & Hoskin D.W. (2020). MDA-MB-231 Breast Cancer Cells Resistant to Pleurocidin-Family Lytic Peptides Are Chemosensitive and Exhibit Reduced Tumor-Forming Capacity. Biomolecules, 10(9), 1220.
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