The U251 cell line was grown in DMEM with high glucose (EuroClone S.p.A., Milano, Italy) supplemented with 10% FBS (EuroClone S.p.A., Milano, Italy), 100 IU/mL penicillin G, 100 μg/mL streptomycin (EuroClone S.p.A., Milano, Italy) in an H2O-saturated 5% CO2 atmosphere at 37 °C.
Dmem with high glucose
Manufactured by Euroclone
Sourced in Italy, United States
DMEM with high glucose is a cell culture medium that provides a nutrient-rich environment for the growth and maintenance of various cell types. It contains a high concentration of glucose, a primary energy source for cells. The formulation is designed to support the optimal growth and proliferation of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Euroclone (3 mentions)
Streptomycin, by Euroclone (1 mentions)
Penicillin g, by Euroclone (1 mentions)
β fgf, by Thermo Fisher Scientific (1 mentions)
Ultra low attachment flask, by Corning (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Euroclone (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Calcium chloride cacl2, by Merck Group (1 mentions)
Minimum essential medium (mem), by Euroclone (1 mentions)
L glutamine, by Euroclone (1 mentions)
3 protocols using dmem with high glucose
1
Culturing U251 Glioma Cells
2
Isolation and Differentiation of Breast Cancer Stem Cells
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BCSCs were isolated from human breast cancer tissues obtained from patients as previously described34 (link) and were cultured in a selective medium34 (link) supplemented with 10 ng/ml βFGF (Peprotech, London, UK), 20 ng/ml EGF (Peprotech) to a final concentration 5 × 104/ml in ultra low attachment flask (Corning, New York, NY, USA) at 37 °C in a 5% (v/v) CO2 humidified chamber. BCSCs were induced to differentiate in order to obtain SDACs by culturing them in adherent condition in D-MEM with high glucose (Euroclone, Milan, Italy) supplemented with 10% (v/v) fetal bovine serum (Euroclone). The tumor s were histopathologically classified as follows: BCSC#1 is an invasive ductal carcinoma, grading G2, ER 90%, PR 60%, HER2/neu 3+ and ki67 >10% BCSC#2 is an invasive ductal carcinoma, grading G2, ER 90%, PR 60%, HER2/neu 3+ and ki67 25%, BCSC#3 is an invasive ductal carcinoma, grading G2, ER 80%, PR 80%, HER2/neu 3+ and ki67 >10%. HER2 status has been assigned according to the FDA guidelines.
3
Culturing and Quiescence of Keratinocytes and Fibroblasts
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Immortalized human keratinocytes (HaCaT) were grown in DMEM with high glucose (Euroclone) supplemented with 10% Fetal Bovine Serum (FBS) (Euroclone), 2% Streptomycin/Penicillin (Euroclone) and 2% L-Glutamine (Euroclone).
LF-1 normal human embryonic lung fibroblasts (kindly provided by J. Sedivy, Brown University, Providence; RI, USA) were grown in MEM (Euroclone) supplemented with 10% FBS (GIBCO), 1% Streptomycin/Penicillin (Euroclone) and 1% L-Glutamine (Euroclone). Cells were cultured in sterility conditions and kept at 37 °C in humidified atmosphere at 5% of CO 2 .
Cell quiescence was obtained by serum starvation: LF-1 fibroblasts reached quiescence within 3 days in medium with 0.5% FBS (23) , while HaCaT keratinocytes required 5 days in medium with 0.1% FBS.
Calcium chloride (CaCl 2 ) (Sigma-Aldrich) was used as supplement in HaCaT cell medium to arrest cell proliferation by inducing cell differentiation (22) . CaCl 2 was added to the culture medium to reach a final 3.8 mM concentration. Cells were grown in CaCl 2 -enriched medium for 5 days.
To evaluate the proliferation state, cells grown on coverslips were incubated for 1 hour in medium containing 20 µM 5-bromo-2'-deoxyuridine (BrdU) or 5-ethynyl-2'-deoxyuridine (EdU), and then cells were fixed in 70% cold ethanol and, if not processed immediately, stored at -20°C.
LF-1 normal human embryonic lung fibroblasts (kindly provided by J. Sedivy, Brown University, Providence; RI, USA) were grown in MEM (Euroclone) supplemented with 10% FBS (GIBCO), 1% Streptomycin/Penicillin (Euroclone) and 1% L-Glutamine (Euroclone). Cells were cultured in sterility conditions and kept at 37 °C in humidified atmosphere at 5% of CO 2 .
Cell quiescence was obtained by serum starvation: LF-1 fibroblasts reached quiescence within 3 days in medium with 0.5% FBS (23) , while HaCaT keratinocytes required 5 days in medium with 0.1% FBS.
Calcium chloride (CaCl 2 ) (Sigma-Aldrich) was used as supplement in HaCaT cell medium to arrest cell proliferation by inducing cell differentiation (22) . CaCl 2 was added to the culture medium to reach a final 3.8 mM concentration. Cells were grown in CaCl 2 -enriched medium for 5 days.
To evaluate the proliferation state, cells grown on coverslips were incubated for 1 hour in medium containing 20 µM 5-bromo-2'-deoxyuridine (BrdU) or 5-ethynyl-2'-deoxyuridine (EdU), and then cells were fixed in 70% cold ethanol and, if not processed immediately, stored at -20°C.
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