Dmem high glucose medium

Manufactured by Corning

Sourced in United States

DMEM high glucose medium is a common cell culture media used to support the growth of a variety of cell types in the laboratory. It provides essential nutrients, amino acids, and other components required for cell proliferation and maintenance. The high glucose concentration in this formulation makes it suitable for culturing cells with high metabolic activity.

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14 protocols using dmem high glucose medium

Cell Culture Conditions for HCT116, U2OS, and HT-1080

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HCT116 (ATCC CCL-247) and U2OS (ATCC HTB-96) cell lines were maintained at 37°C and 5% CO₂ in DMEM high glucose (GE Healthcare) supplemented with 10% FBS (Atlanta Biologicals), 2mM L-glutamine (Gemini Biosciences), 1mM sodium pyruvate (Gibco), 1x MEM non-essential amino acids solution (Gibco), 40 U/mL penicillin and 40 μg/mL streptomycin (Gemini Biosciences).
HT-1080 (ATCC CCL-121) cells were grown in DMEM high glucose medium (Corning Life Science) supplemented with 1% non-essential amino acids (Life Technologies). HT-1080 cells used in this study were stably infected with Nuc:mKate2, a red fluorescent protein targeted to the nucleus.^{56 (link)}

Broder Schmidt H., Jaafar Z.A., Erik Wulff B., Rodencal J.J., Hong K., Aziz-Zanjani M.O., Jackson P.K., Leonetti M.D., Dixon S.J., Rohatgi R, & Brandman O. (2022). Oxaliplatin disrupts nucleolar function through biophysical disintegration. Cell reports, 41(6), 111629.

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Cell Culture Protocols for Endometrial and Cancer Cell Lines

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ECSCs were cultured in serum-free medium, DMEM/F12 (1:1) (Corning, New York, USA) containing 2% B27 Supplement (Gibco, New York, USA), 20 ng/mL EGF (PeproTech, New Jersey, USA), 20 ng/mL bFGF (PeproTech), and 1% penicillin–streptomycin (Invitrogen, Carlsbad, California, USA). Ishikawa cell line (Shanghai huiying, Shanghai, China) and HEC-1A cell line (Genechem, Shanghai, China) were cultured in α-MEM medium (Bioind, Kibbutz Beit Haemek, Israel) and McCoy's 5A medium (Bioind), respectively. The medium contained 10% fetal bovine serum (FBS) (Bioind) and 1% penicillin–streptomycin (Invitrogen). HEK293T cells were cultured in DMEM/high-glucose medium (Corning). The medium contained 10% FBS (Bioind) and 1% penicillin–streptomycin (Invitrogen). All cells were cultured in a humidified incubator at 37° C with 5% CO₂.

Li Y., Huo J., He J, & Ma X. (2021). LncRNA MONC suppresses the malignant phenotype of Endometrial Cancer Stem Cells and Endometrial Carcinoma Cells by regulating the MiR-636/GLCE axis. Cancer Cell International, 21, 331.

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Endogenous Co-IP of RBP Complexes

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HepG2 cells were grown in DMEM high glucose medium (Corning) with 10% fetal bovine serum (FBS) (GIBCO). K562 cells were maintained in RPMI 1640 (Corning) with 10% FBS. Endogenous Co-IP was carried out using indicated antibody and Protein A/G Magnetic Beads (Thermofisher, 88803). After RNase A/T1 Mix (Thermofisher, EN0551) treatment and several times of washes, precipitated protein were eluted in SDS loading buffer and separated by SDS-PAGE, transferred onto PVDF membranes (Millipore) and detected blots with appropriate antibodies. Antibodies used in this study include: anti-SRSF1 (Abcam, ab38017), anti-FKBP4 (Abcam, ab129097), anti-EIF4G2 (Cell Signaling, 5169), and anti-IGF2BP1 (Abcam, ab107205).

Li Y.E., Xiao M., Shi B., Yang Y.C., Wang D., Wang F., Marcia M, & Lu Z.J. (2017). Identification of high-confidence RNA regulatory elements by combinatorial classification of RNA–protein binding sites. Genome Biology, 18, 169.

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Cell Culture Conditions for HCT116, U2OS, and HT-1080

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Isolation and Culture of Muse Cells

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MECs were suspended in DMEM high glucose medium (Corning, NY, USA) supplemented with 0.9% MethoCult (H4100, StemCell Technologies Inc.), 10% FBS (Gibco, Grand Island, NY), and 1% penicillin/streptomycin (Gibco). Then, the mixture was seeded in poly-HEMA (P3932, Sigma–Aldrich, Saint Louis, MO, USA) pre-treated 96-well cell culture plate by limited dilution method, 70 μL for each well. Wells containing 1–3 cells were chosen for following studies. 30 μL complete DMEM medium containing 10% FBS and 1% penicillin/streptomycin was added every 3 days. After 7–10 days, single Muse cell will grow to a Muse-cluster (M-cluster), while non-Muse cell cannot generate a cluster. The generated M-clusters were used for immunocytochemistry, ALP staining and tumorigenicity assays. To prepare adherent Muse cells, M-clusters were collected by centrifuging at 1500 rpm for 5 min, resuspended in complete medium, and then placed in cell culture dishes for normal adherent culture. When reaching 70%–80% confluency, cells were used for multilineage differentiation, immunocytochemistry, migration assay in vitro and cell transplantation in vivo.

Li H., Wei J., Li M., Li Y., Zhang T., Tian J., Liu X., Li K, & Lin J. (2024). Biological characteristics of Muse cells derived from MenSCs and their application in acute liver injury and intracerebral hemorrhage diseases. Regenerative Therapy, 27, 48-62.

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Neonatal Fibroblast Senescence Modeling

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Neonatal human foreskin fibroblasts (HFFs) from the ATCC were maintained in DMEM high-glucose medium (Corning) supplemented with 10% HyClone fetal bovine serum (Thermo Scientific) at 37°C under 5% CO₂. In the TIS model, cells were seeded at 1×10³/cm² in 10-cm culture dishes. Twenty-four hours after seeding, cells were incubated in complete medium with or without 20 μM ICRF-187 (Selleck Chemicals) for 24h and then the media was changed. Cells were incubated in complete medium with or without 100 ng/ml doxorubicin (Sigma-Aldrich) for 12 h, cultured in fresh complete medium and recorded as day 0. Cells were then prepared for experiments with regular medium changes at different time points. RS was induced by serial passaging of HFFs at a ratio of 1:3 and seeded at ~6 000 cells/cm² in 10-cm dishes every 4-7 days.

Wang D., Liu Y., Zhang R., Zhang F., Sui W., Chen L., Zheng R., Chen X., Wen F., Ouyang H.W, & Ji J. (2016). Apoptotic transition of senescent cells accompanied with mitochondrial hyper-function. Oncotarget, 7(19), 28286-28300.

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Culturing Human Breast Cancer Cells

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Human breast cancer cell lines, MDA-MB-231 harboring oncogenic KRAS mutation and MDA-MB-468 with wild-type KRAS, were purchased from American Type Culture Collection. DMEM/high glucose medium (Corning) supplemented with 10% fetal bovine serum (Gibco) and 100 U/mL penicillin-streptomycin (Gibco) was used to maintain both cell lines. Cells were kept in a humidified atmosphere with 5% carbon dioxide at 37°C.

Liu X, & Ghosh D. (2019). Intracellular nanoparticle delivery by oncogenic KRAS-mediated macropinocytosis. International Journal of Nanomedicine, 14, 6589-6600.

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Culturing MDA-MB-231, IGROV1, and UC-MSCs

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MDA-MB-231 human breast cancer cells and IGROV1 human ovarian cancer cells were cultured with DMEM (high glucose) medium (Corning, Lowell, MA) supplemented with 10% fetal bovine serum (FBS) (Corning, Lowell, MA) and 1% penicillin streptomycin solution (Gibco, Rockville, MD). Medium for MDA-MB-231 cells was also supplemented with 1% MEM nonessential amino acid solution (NEAA; Gibco). UC-MSCs were isolated as described before [17 (link), 18 (link)] and cultured with DMEM/F12 medium (Gibco) containing 10% FBS (Corning), 1% penicillin streptomycin solution (Gibco), and 10 ng/ml human recombinant epidermal growth factor (EGF; Gibco). All cell lines were maintained at 37°C in a 5% CO2 incubator. To be trackable in direct coculturing model, MDA-MB-231 cells were transduced with lentiviral vector carrying green fluorescence protein (GFP) and selected with blasticidin.

Luo X., Huang S., He N., Liu C., Chen Y., Liu Y., Mi X., Li N., Sun P., Li Z., Xiang R, & Su W. (2018). Inflammatory Human Umbilical Cord-Derived Mesenchymal Stem Cells Promote Stem Cell-Like Characteristics of Cancer Cells in an IL-1β-Dependent Manner. BioMed Research International, 2018, 7096707.

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Culturing Cholangiocarcinoma Cell Lines

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The 4 cholangiocarcinoma cell lines (HCCC-9810, HUCCT1, QBC939, and RBE cells) and human intrahepatic biliary epithelial cell line (HIBEC cells) used in this study were all from BeNa Culture Collection (China). HCCC-9810, HUCCT1 and HIBEC cells were grown in RPMI-1640 medium (Corning) containing 10% foetal bovine serum (FBS, Ausbian). QBC939 and RBE cells were grown in DMEM high glucose medium (Corning) containing 10% FBS. All cells were cultured at 37 °C in an incubator with 5% CO₂.

Zhang C., Wang Y., Wu G., Sun N., Bai H., Li X., Han S., Zhou H., Qi R, & Zhang J. (2024). RPL35A promotes the progression of cholangiocarcinoma by mediating HSPA8 ubiquitination. Biology Direct, 19, 16.

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Culturing Human Breast Cancer Cell Lines

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MDA-MB-231 and SK-BR-3 human breast cancer cell lines (purchased from Shanghai Jikeji Company) were cultured in 37°C constant temperature incubator (containing 5% CO2) [Sanyo Sanyo, Mco-175] with complete medium prepared with DMEM (high glucose) medium (Corning, USA) + 10% fetal bovine serum [FBS (Corning, USA)] + 1% di-anti-penicillin/streptomycin [P/S (Corning, USA)]. HCC1937, T47D and MCF7 human breast cancer cell lines (purchased from Shanghai Jikaine Company) were cultured in a 37°C constant temperature incubator (containing 5% CO2) [Sanyo Sanyo, Mco-175] in a complete medium with RPMI1640 medium (Corning, USA) + 10% fetal bovine serum (FBS) + 1% double-resistant penicillin/streptomycin (P/S).

Yu P., Zhu L., Cui K., Du Y., Zhang C., Ma W, & Guo J. (2021). B4GALNT2 Gene Promotes Proliferation, and Invasiveness and Migration Abilities of Model Triple Negative Breast Cancer (TNBC) Cells by Interacting With HLA-B Protein. Frontiers in Oncology, 11, 722828.

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