Human umbilical vein endothelial cells (HUVECs) and human pulmonary microvascular endothelial cells (HPMECs) were purchased from Typical Animal Reserve Center of China (Shanghai, China) and ScienCell (Carlsbad, CA), respectively. Both HUVECs and HPMECs were cultured in endothelial cell medium (ECM, ScienCell, Carlsbad, CA) and cells harvested at passages 3–10 were used in assays. Human hepatocellular carcinoma (HCC) Hep3B cells were obtained from ATCC and cultured in DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin. Human lung carcinoma cells (A549) were obtained from the China Centre for Type Culture Collection (CCTCC, Wuhan, China) and cultured in RPMI-1640 supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin. Human erythroleukemia TF-1 cells were purchased from ATCC and cultured in RPMI-1640 supplemented with 2 ng/mL rhGM-CSF and 10% FBS. Human glioblastoma U87-MG cells were from ATCC (Molsheim, France) and cultured in DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin. All the above cells were incubated at 37 °C under 95% air and 5% CO2. A hypoxic condition was created in a hypoxia chamber (Billups-Rothenberg, Inc., San Diego, CA) equilibrated with certified gas containing 1% O2, 5% CO2, and 94% N2.
U87mg cells
Manufactured by American Type Culture Collection
Sourced in United States
U87MG cells are a well-characterized and widely used glioblastoma cell line derived from human brain tissue. They are suitable for a variety of cell biology and cancer research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (17 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Dmem f12, by Merck Group (2 mentions)
Huvecs, by Lonza (2 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (2 mentions)
B16f10 cells, by American Type Culture Collection (2 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (2 mentions)
Hep3b cells, by American Type Culture Collection (1 mentions)
Huvecs, by ScienCell (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Hpmec, by ScienCell (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Gamma counter, by PerkinElmer (1 mentions)
61 protocols using u87mg cells
1
Endothelial and Cancer Cell Culture Conditions
2
Culturing Murine and Human Cell Lines
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Murine astrocytoma ALTS1C1 cells (BCRC60582, Taiwan) [22 (link)], murine pancreatic UN-KC6141 cells (kindly provided by Prof. Batra, S. K. from Department of Pathology and Microbiology, University of Nebraska Medical Center, USA) [23 (link)], and human glioma U87-MG cells (ATCC No. CRL-9589, Manassas, VA, USA) were maintained in Dulbecco’s modified Eagle’s medium (GIBCO, Thermo Fisher Scientific, Inc., USA) with 10% fetal bovine serum (GIBCO), and 1% penicillin–streptomycin (GIBCO) at 37 ° C in a humidified 5% CO2/air atmosphere.
3
Culturing U87-MG Cells Under Hypoxia
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U87-MG cells were newly purchased from ATCC. Cells were routinely cultured in DMEM medium, supplemented with 10% foetal bovine serum (FBS), 2 mM L-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin (growth medium). All cells were grown in humidified 5% CO2 incubator at 37 °C. For hypoxia experiments, cells were incubated in humidified Sci-tive NN Hypoxia workstation (Ruskinn Technology) set at 5% CO2, 1% O2, and 37 °C.
4
Cell Culture Protocols for Cancer Research
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CHO-K1 (ECACC, Wiltshire, UK) and CHO-ETBR cells (CHO-K1 cells stably transfected with vector expressing human ETBR [40] (link)) were donated by the laboratory of Frédéric Ducancel (CEA, Saclay, France). U87-MG cells (ATCC, Manassas, VA) were donated by the laboratory of Andreas Jacobs (Max Plank Institute, Köln, Germany). 4T1 and EMT6 cells (ATCC, Manassas, VA) were donated by the laboratory of Lina Bolotine (Research Centre for Automatic Control (CRAN), Nancy-University, UMR CNRS, France). MDA-MB-231, A-431, MCF-7 and PC3 cells were purchased from ATCC (Manassas, VA).
CHO cells were grown in Nutrient Mixture F-12 HAM supplemented with 10% Foetal Bovin Serum (FBS). MCF-7 cells were grown in RPMI media supplemented with 10% FBS. A431, MDA-MB-231, U87-MG, 4T1, and EMT6 cells were grown in DMEM media with 10% FBS. All the cell types were grown at 37°C in a 5% CO2 atmosphere.
CHO cells were grown in Nutrient Mixture F-12 HAM supplemented with 10% Foetal Bovin Serum (FBS). MCF-7 cells were grown in RPMI media supplemented with 10% FBS. A431, MDA-MB-231, U87-MG, 4T1, and EMT6 cells were grown in DMEM media with 10% FBS. All the cell types were grown at 37°C in a 5% CO2 atmosphere.
5
Cellular Uptake Assay for 68Ga-RGD Peptides
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Human glioblastoma multiforme U-87 MG cells (ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle Medium (Sigma Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum, 0.1 mM non-essential amino acids and 1.0 mM sodium pyruvate at 37 °C in a 5% carbon dioxide humidified incubator. The cells were subcultured and used for experiments at a confluency of 70–90%.
Cell uptake assay was performed in 24-well plates with U-87 MG cell at high confluency. The assay itself was carried out using a dedicated buffer instead of the regular cell culture media. The buffer consists of 25 mM Tris/HCl, 5.4 mM KCl, 1.8 mM CaCl2, 0.8 mM MgSO4, 5 mM glucose, 140 mM NaCl in H2O [34 (link)].
The cells were incubated with 68Ga-labeled RGD peptides (20 µM) alone and also with excess (2 mM) of cold RGD peptide for 90 min (37 °C) in the buffer described above. The cells were rinsed with PBS, lysed in 0.1 M NaOH and measured for radioactivity in a gamma counter (PerkinElmer, Waltham, Massachusetts, USA). The uptake was calculated as the mean of the percentage of the applied dose ± standard deviation.
Cell uptake assay was performed in 24-well plates with U-87 MG cell at high confluency. The assay itself was carried out using a dedicated buffer instead of the regular cell culture media. The buffer consists of 25 mM Tris/HCl, 5.4 mM KCl, 1.8 mM CaCl2, 0.8 mM MgSO4, 5 mM glucose, 140 mM NaCl in H2O [34 (link)].
The cells were incubated with 68Ga-labeled RGD peptides (20 µM) alone and also with excess (2 mM) of cold RGD peptide for 90 min (37 °C) in the buffer described above. The cells were rinsed with PBS, lysed in 0.1 M NaOH and measured for radioactivity in a gamma counter (PerkinElmer, Waltham, Massachusetts, USA). The uptake was calculated as the mean of the percentage of the applied dose ± standard deviation.
6
Cell Culture Protocols for Cancer Research
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U87-MG cells were purchased from ATCC (Manassas, VA). U251-MG cells were purchased from Sigma-Aldrich. U87-MG and U251-MG were passaged regularly and cultured with DMEM (Corning Life Sciences, Corning, NY) in the presence of 10% v/v fetal bovine serum (Atlanta Biologicals; Life Technologies). Immortalized mammary epithelial cells (IMECs) were a kind gift from Dr. James DiRenzo. IMECs were cultured in DMEM/F12 50/50 media supplemented with 5% FBS, 2 mM glutamine (Gibco), 5 μg/mL insulin (Akron Biotech), 500 μg/mL hydrocortisone (MP Biomedical), and 10 ng/mL recombinant human epidermal growth factor (Promega). Cells were routinely verified as mycoplasma free with the MycoProbe kit (R&D Systems).
7
Culturing Highly Invasive U87 MG Glioblastoma Cells
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Human glioblastoma U87 MG cells (ATCC) were used for all experiments because of their highly invasive nature. Cells were cultured at 37 °C in a 5% CO2 atmosphere in DMEM/F12 (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Fisher Scientific, Hampton, NH, USA), 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA) and 1% MycoZap (Life Technologies, Carlsbad, CA, USA). Cells were fed 2–3 times a week and passaged at confluence prior to use.
8
Establishing Ba/F3 and U87MG Cell Lines
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Ba/F3 cells were purchased from DSMZ (Braunschweig, Germany) and maintained in RPMI1640 medium (Life Technologies) supplemented with 10% FBS, 1% l -glutamine, and 10 ng/mL mouse IL3 (R&D Systems). Lack of endogenous murine erbB family member protein expression in mammalian hematopoietic cells (17 (link)) was confirmed in parental Ba/F3 cells (Supplementary Fig. S1). Ba/F3 transformed cells were maintained in the medium with 15-µg/mL blasticidin S HCl (Life Technologies). U87MG cells were purchased from ATCC and maintained in MEM (Life Technologies) supplemented with 10% FBS and 1% l -glutamine. The cell lines were periodically checked for Mycoplasma using MycoAlert Plus Mycoplasma Detection Kit (Lonza).
9
Umbilical Cord Blood and Peripheral Blood Protocol
10
Culturing Primary Astrocytes and Glioma Cells
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