Dulbecco s modified eagle s medium high glucose medium

Manufactured by Thermo Fisher Scientific

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Dulbecco's modified Eagle's medium/high glucose medium is a cell culture medium formulation developed by Dulbecco and Freeman. It is commonly used to support the growth of a variety of mammalian cell types in vitro. The medium provides essential nutrients, salts, and a high concentration of glucose to sustain cell proliferation and viability.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Poly d lysine, by Merck Group (2 mentions) Penicillin, by Quality Biological (2 mentions) Streptomycin, by Quality Biological (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by AppliChem (1 mentions) Tc 100 medium, by AppliChem (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Q vd oph, by Merck Group (1 mentions) Z vad fmk, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) X tremegene hp dna transfection reagent, by Roche (1 mentions) Calf bovine serum, by Thermo Fisher Scientific (1 mentions) Victor2 multilabel plate reader, by PerkinElmer (1 mentions) Penicillin streptomycin, by Nacalai Tesque (1 mentions) Bz x710 fluorescence microscope, by Keyence (1 mentions) Adipored assay reagent, by Lonza (1 mentions) Hanks balanced salt solution without ca2 and mg2, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

10 protocols using dulbecco s modified eagle s medium high glucose medium

Silkworm Strain Characterization and Cell Line Maintenance

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A bivoltine silkworm strain, u11, which shows minute wing (mw) phenotype, was used as the natural mutant strain. Another bivoltine silkworm strain, p50, which shows normal wings, was used as wild type animals for the back‐crossing strategy. A multivoltine silkworm strain, Nistari, was used in generating CRISPR/Cas9‐mediated mutagenesis and subsequent experiments. All silkworm strains were preserved in the Sericultural Research Institute, Chinese Academy of Agricultural Sciences, and larvae were fed with fresh mulberry leaves at 25 °C under standard conditions (Tan et al., 2005).
The BmN cell line originating from the B. mori ovary was used (Pan et al., 2010). The cells were maintained in TC‐100 medium (AppliChem, Gatersleben, Germany) containing 10% fetal bovine serum in 25 cm² Petri dishes at 27 °C. The mammalian HEK293T cell line was maintained at 37 °C under 5% CO₂ in Dulbecco's modified Eagle's medium high glucose medium (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (Gibco).

Yu Y., Liu X., Ma X., Zhang Z., Wang T., Sun F., Hou C, & Li M. (2018). A palmitoyltransferase Approximated gene Bm‐app regulates wing development in Bombyx mori. Insect Science, 27(1), 2-13.

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Apoptosis Induction in Cell Lines

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BMK WT or bax/bak DKO (provided by the Eileen White lab, Rutgers Cancer Institute of New Jersey), HeLa, and Cos-7 cell lines were grown under standard cell culture conditions (37°C, 5% CO₂) in Dulbecco’s modified Eagle’s medium high-glucose medium (Gibco) supplemented with 10% fetal bovine serum (Sigma-Aldrich), penicillin (100 U/ml), and streptomycin (100 μg/ml) (Gibco). Cell transfection was performed using X-tremeGENE HP DNA Transfection Reagent (Roche) according to the manufacturer’s protocol. Six hours after transfection, cell medium was removed, and cells were incubated for 24 hours with a new medium. When required, cell detachment was prevented using Z-VAD-FMK (10 to 20 μM; Sigma-Aldrich) and Q-VD-Oph (10 μM; Sigma-Aldrich), which was added before the transfection or 6 hours after transfection. Alternatively and when stated, trBax cells were cotransfected with baculovirus caspase inhibitor p35.

Popgeorgiev N., Sa J.D., Jabbour L., Banjara S., Nguyen T.T., Akhavan-E-Sabet A., Gadet R., Ralchev N., Manon S., Hinds M.G., Osigus H.J., Schierwater B., Humbert P.O., Rimokh R., Gillet G, & Kvansakul M. (2020). Ancient and conserved functional interplay between Bcl-2 family proteins in the mitochondrial pathway of apoptosis. Science Advances, 6(40), eabc4149.

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Quantifying Adipocyte Differentiation in 3T3-L1 Cells

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Mouse 3T3‐L1 preadipocytes were purchased from DS Pharma Biomedical. Preadipocytes were cultured in Dulbecco's modified Eagle's medium–high glucose medium (Gibco, Gaithersburg, MD, USA) supplemented with 10% calf bovine serum (Gibco) and penicillin–streptomycin (Nacalai Tesque) at 37°C in a humidified 5% CO₂ atmosphere until confluence was reached in a 96‐well plate format. Two days after confluence (day 0), cells were stimulated to differentiate by culturing in adipocyte differentiation medium (ADM; DS Pharma Biomedical) for 3 days. Cells were then maintained in adipocyte maintenance medium (DS Pharma Biomedical) for an additional 4 days. Extracts or fractions (10 μg/ml) were administered from day 0 of adipocyte differentiation. On day 7, intracellular lipid droplets were stained using AdipoRed Assay Reagent (Lonza) according to the manufacturer's instructions. After obtaining images using a BZ‐X710 fluorescence microscope (Keyence), intracellular lipid accumulation was quantified by measuring fluorescence (Ex 485 nm/Em 590 nm) using a Victor2 multilabel plate reader (PerkinElmer).

Matsuoka I., Hata K., Katsuzaki H., Nakayama H., Zang L., Ota M., Kim Y., Chu D., Juneja L.R., Nishimura N, & Shimada Y. (2022). Zebrafish obesogenic test identifies anti‐adipogenic fraction in Moringa oreifera leaf extracts. Food Science & Nutrition, 10(4), 1248-1256.

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Cell Culture Maintenance Protocols

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All the cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and were routinely maintained in Dulbecco's modified Eagle's medium/high glucose medium (Gibco-BRL, Rockville, IN, USA) with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin in a humidified incubator under the conditions of 5% CO₂ at 37 °C.

Liang Y., Song X., Li Y., Sang Y., Zhang N., Zhang H., Liu Y., Duan Y., Chen B., Guo R., Zhao W., Wang L, & Yang Q. (2018). A novel long non-coding RNA-PRLB acts as a tumor promoter through regulating miR-4766-5p/SIRT1 axis in breast cancer. Cell Death & Disease, 9(5), 563.

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Culturing HEK-293T and DRG Neurons for Viral Transduction

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HEK-293T cell cultures and DRG neuronal cultures were prepared according to previously described methods (10 (link), 32 (link)). Briefly, HEK-293T cells were cultured in Dulbecco’s modified Eagle’s medium/high glucose medium (Gibco/Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS) and 1% antibiotics. DRGs from adult mice were collected in ice-cold Neurobasal medium (Life Technologies) with 10% FBS (J R Scientific) and penicillin (100 U/ml) and streptomycin (100 µg/ml) (Quality Biological) and then digested with enzyme solution [dispase (5 mg/ml) and collagenase type I (1 mg/ml)] in Hanks’ balanced salt solution without Ca²⁺ and Mg²⁺ (Life Technologies) at 37°C for 20 to 25 min. After trituration and centrifugation, the dissociated neurons were resuspended in mixed Neurobasal medium and plated into six-well plates coated with poly-d-lysine (50 µg/ml) (Sigma) at a density of 1.5 × 10⁵ to 4 × 10⁵ cells. The neurons were incubated in a humidified 95% O₂ and 5% CO₂ atmosphere at 37°C. For viral infection, 2 µl of AAV5 virus (titer ≥ 1 × 10¹²/ml) was added to each 2-ml well after 24 hours of incubation. For siRNA transfection, siRNA was diluted to the concentration of 100 nM and transfected to cultured cells by Lipofectamine 2000 (Invitrogen), according to the manufacturer’s instruction. Cells were collected 2 or 3 days later for Western blot analysis.

Li Z., Mao Y., Liang L., Wu S., Yuan J., Mo K., Cai W., Mao Q., Cao J., Bekker A., Zhang W, & Tao Y.X. (2017). The transcription factor C/EBPβ in the dorsal root ganglion contributes to peripheral nerve trauma–induced nociceptive hypersensitivity. Science signaling, 10(487), eaam5345.

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Primary DRG Neuron and HEK-293T Cell Culture

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HEK‐293T cell cultures and DRG neuronal cultures were prepared according to previously described methods.^[¹⁰ , ¹⁸ (link), ⁴⁹ (link), ⁵⁴ (link)
^] Briefly, HEK‐293T cells were cultured in Dulbecco's modified Eagle's medium/high glucose medium (Gibco/Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS) and 1% antibiotics. For primary DRG neuronal cultures, after 4‐week‐old CD1 mice were euthanized with isoflurane, all DRGs were collected in cold Neurobasal Medium (Gibco/ThermoFisher Scientific) containing 10% FBS (JR Scientific, Woodland, CA), 100 units mL⁻¹ Penicillin and 100 µg mL⁻¹ Streptomycin (Quality Biological, Gaithersburg, MD). The DRGs were then treated with enzyme solution (5 mg mL⁻¹ dispase, 1 mg mL⁻¹ collagenase type I in Hanks’ balanced salt solution (HBSS) without Ca²⁺ and Mg²⁺ (Gibco/ThermoFisher Scientific). After trituration and centrifugation, dissociated cells were resuspended in a mixed Neurobasal Medium and plated in a six‐well plate coated with 50 µg mL⁻¹ poly‐D‐lysine (Sigma, St. Louis, MO). The cells were incubated at 95% O₂, 5% CO₂, and 37 ℃. On the second day, 2–10 µL of virus (titer ≥1 × 10¹²/µL) or siRNA (100 x 10^–9m; transfected with Lipofectamine 2000) was added to every 2 mL well. Cells were collected 3 days later.

Pan Z., Du S., Wang K., Guo X., Mao Q., Feng X., Huang L., Wu S., Hou B., Chang Y., Liu T., Chen T., Li H., Bachmann T., Bekker A., Hu H, & Tao Y. (2021). Downregulation of a Dorsal Root Ganglion‐Specifically Enriched Long Noncoding RNA is Required for Neuropathic Pain by Negatively Regulating RALY‐Triggered Ehmt2 Expression. Advanced Science, 8(13), 2004515.

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Culturing HepG2 Liver Cancer Cells

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The HepG2 HCC cell lines were obtained from KCLB (Korean Cell Line Bank, Seoul, Korea). HepG2 cells were maintained in Dulbecco's modified Eagle's medium‒high glucose medium (Thermo Fisher Scientific, Carlsbad, CA). The medium was supplemented with 10% fetal bovine serum (GibcoBRL, Carlsbad, CA) and 1% antibiotics (Thermo Fisher Scientific) at 37°C in a humidified atmosphere with 5% CO₂ in an incubator.

Lee S.C., Kim K.H., Kim O.H., Lee S.K., Hong H.E., Choi B.J., Jeong W, & Kim S.J. (2017). Everolimus Plus Ku0063794 Regimen Promotes Anticancer Effects against Hepatocellular Carcinoma Cells through the Paradoxical Inhibition of Autophagy. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 50(3), 1023-1038.

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Cultivation of Human Colon Cancer Cells

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The human colon cancer cell line HCA-7 colony 29 (European Collection of Cell Cultures, Salisbury, UK) was grown in Dulbecco׳s modified Eagle׳s medium high-glucose medium (Thermo Fisher Scientific, UT, USA) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, UT, USA) and cultured in an incubator containing a 95% humidified atmosphere with 5% CO₂ at 37 °C. Cells were sub-cultured at a ratio of 1:5–1:8 after they had reached ~90% confluency.

Xu Y., Qi J., Yang X., Wu E, & Qian S.Y. (2014). Free radical derivatives formed from cyclooxygenase-catalyzed dihomo-γ-linolenic acid peroxidation can attenuate colon cancer cell growth and enhance 5-fluorouracil׳s cytotoxicity. Redox Biology, 2, 610-618.

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Transient Expression of SOD1 in HEK293T Cells

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HEK293T cells (CRL-3216; American Type Culture Collection, Manassas, VA) were maintained in Dulbecco’s modified Eagle’s medium high-glucose medium (Life Technologies, Carlsbad, CA) supplemented with 2 mmol dm⁻³ L-glutamine (Life Technologies), 100 U/mL penicillin and streptomycin (Life Technologies), and 10% fetal bovine serum (FBS; Gibco-Thermo Fisher Scientific, Waltham, MA, USA) in uncoated 75 cm² plastic flasks and incubated at 37°C, 5% CO₂ in a humidified atmosphere. The cells were transiently transfected with the pHLsec plasmid (28 (link)) containing the complementary DNA of human superoxide dismutase 1 (SOD1) (UniProtKB: P00441) using branched polyethylenimine ((PEI) average Mw 25 kDa; Sigma-Aldrich, St. Louis, MO), as previously described (14 (link)). A 1:2 DNA:PEI weight ratio (25 μg SOD1 DNA, 50 μg PEI per 75 cm² flask) was used. Transient expression of SOD1 was carried out for 48 h in [U-¹⁵N]-BioExpress6000 medium (Cambridge Isotope Laboratories, Tewksbury, MA) supplemented with 2% FBS, 100 U/mL penicillin/streptomycin, and 10 μM ZnSO₄.

Cerofolini L., Giuntini S., Barbieri L., Pennestri M., Codina A., Fragai M., Banci L., Luchinat E, & Ravera E. (2018). Real-Time Insights into Biological Events: In-Cell Processes and Protein-Ligand Interactions. Biophysical Journal, 116(2), 239-247.

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Culturing OSCC and Normal Oral Keratinocytes

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The human OSCC cell line CAL27 was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA), and cultured in Dulbecco's modified Eagle's medium high-glucose medium (Life Technologies, Grand Island, NY, USA) supplemented with heat-inactivated 10% fetal bovine serum (HyClone Laboratories, Logan, UT, USA). The normal oral keratinocyte NOK-SI cell line was kindly provided by the National Institute of Health (Bethesda, MD, USA) and was grown in keratinocyte serum-free medium containing human recombinant epidermal growth factor and bovine pituitary extract (Life Technologies). Cells were cultured at 37°C in a humidified atmosphere of 5% CO₂ and 95% air.

ZENG Q., TAO X., HUANG F., WU T., WANG J., JIANG X., KUANG Z, & CHENG B. (2016). Overexpression of miR-155 promotes the proliferation and invasion of oral squamous carcinoma cells by regulating BCL6/cyclin D2. International Journal of Molecular Medicine, 37(5), 1274-1280.

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